IL-8 receptor antagonists

ABSTRACT

Novel IL-8 receptor antagonists and methods of using them are provided.

This application is a 371 of PCT/US97/10900 filed Jun. 24, 1997 andclaims the benefit of Provisional Application No. 60/020,655 filed Jun.27, 1996.

FIELD OF THE INVENTION

This invention relates to a novel group phenyl urea compounds, processesfor the preparation thereof, the use thereof in treating IL-8, GROα,GROβ, GROγ, NAP-2, and ENA-78 mediated diseases and pharmaceuticalcompositions for use in such therapy.

BACKGROUND OF THE INVENTION

Many different names have been applied to Interleukin-8 (IL-8), such asneutrophil attractant/activation protein-1 (NAP-1), monocyte derivedneutrophil chemotactic factor (MDNCF), neutrophil activating factor(NAF), and T-cell lymphocyte chemotactic factor. Interleukin-8 is achemoattractant for neutrophils, basophils, and a subset of T-cells. Itis produced by a majority of nucleated cells including macrophages,fibroblasts, endothelial and epithelial cells exposed to TNF, IL-1α,IL-1β or LPS, and by neutrophils themselves when exposed to LPS orchemotactic factors such as FMLP. M. Baggiolini et al, J. Clin. Invest.84, 1045 (1989); J. Schroder et al, J. Immunol. 139, 3474 (1987) and J.Immunol. 144, 2223 (1990); Strieter, et al, Science 243, 1467 (1989) andJ. Biol. Chem. 264, 10621 (1989); Cassatella et al, J. Immunol. 148,3216 (1992).

Groα, GROβ, GROγ and NAP-2 also belong to the chemokine α family. LikeIL-8 these chemokines have also been referred to by different names. Forinstance GROα, β, γ have been referred to as MGSAα, β and γ respectively(Melanoma Growth Stimulating Activity), see Richmond et al, J. CellPhysiology 129, 375 (1986) and Chang et al, J. Immunol 148, 451 (1992).All of the chemokines of the α-family which possess the ELR motifdirectly preceding the CXC motif bind to the IL-8 B receptor.

IL-8, Groα, GROβ, GROγ, NAP-2 and ENA-78 stimulate a number of functionsin vitro. They have all been shown to have chemoattractant propertiesfor neutrophils, while IL-8 and GROα have demonstrated T-lymphocytes,and basophiles chemotactic activity. In addition IL-8 can inducehistamine release from basophils from both normal and atopic individualsGRO-α and IL-8 can in addition, induce lysozomal enzyme release andrespiratory burst from neutrophils. IL-8 has also been shown to increasethe surface expression of Mac-1 (CD11b/CD18) on neutrophils without denovo protein synthesis. This may contribute to increased adhesion of theneutrophils to vascular endothelial cells. Many known diseases arecharacterized by massive neutrophil infiltration. As IL-8, Groα, GROβ,GROγ and NAP-2 promote the accumulation and activation of neutrophils,these chemokines have been implicated in a wide range of acute andchronic inflammatory disorders including psoriasis and rheumatoidarthritis, Baggiolini et al, FEBS Lett. 307, 97 (1992); Miller et al,Crit. Rev. Immunol. 12, 17 (1992); Oppenheim et al, Annu. Rev. Immunol.9, 617 (1991); Seitz et al., J. Clin. Invest. 87, 463 (1991); Miller etal., Am. Rev. Respir. Dis. 146, 427 (1992); Donnely et al., Lancet 341,643 (1993). In addition the ELR chemokines (those containing the aminoacids ELR motif just prior to the CXC motif) have also been implicatedin angiostasis. Strieter et al, Science 258, 1798 (1992).

In vitro, IL-8, Groα, GROβ, GROγ, and NAP-2 induce neutrophil shapechange, chemotaxis, granule release, and respiratory burst, by bindingto and activating receptors of the seven-transmembrane, G-protein-linkedfamily, in particular by binding to IL-8 receptors, most notably theB-receptor. Thomas et al., J. Biol. Chem. 266, 14839(1991); and Holmeset al., Science 253, 1278 (1991). The development of non-peptide smallmolecule antagonists for members of this receptor family has precedent.For a review see R. Freidinger in: Progress in Drug Research, Vol. 40,pp. 33-98, Birkhauser Verlag, Basel 1993. Hence, the IL-8 receptorrepresents a promising target for the development of novelanti-inflammatory agents.

Two high affinity human IL-8 receptors (77% homology) have beencharacterized: IL-8Rα, which binds only IL-8 with high affinity, andIL-8Rβ, which has high affinity for IL-8 as well as for GRO-α, GROβ,GROγ and NAP-2. See Holmes et al., supra; Murphy et al., Science 253,1280 (1991); Lee et al., J. Biol. Chem. 267, 16283 (1992); LaRosa etal., J. Biol. Chem. 267, 25402 (1992); and Gayle et al., J. Biol. Chem.268,7283 (1993).

There remains a need for treatment, in this field, for compounds whichare capable of binding to the IL-8 α or β receptor. Therefore,conditions associated with an increase in IL-8 production (which isresponsible for chemotaxis of neutrophil and T-cells subsets into theinflammatory site) would benefit by compounds which are inhibitors ofIL-8 receptor binding.

SUMMARY OF THE INVENTION

This invention provides for a method of treating a chemokine mediateddisease, wherein the chemokine is one which binds to an IL-8 α or βreceptor and which method comprises administering an effective amount ofa compound of Formula (I) or a pharmaceutically acceptable salt thereof.In particular the chemokine is IL-8.

This invention also relates to a method of inhibiting the binding ofIL-8 to its receptors in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (I).

Compounds of Formula (I) useful in the present invention are representedby the structure:

wherein

X is oxygen or sulfur;

R is any functional moiety having an ionizable hydrogen and a pKa of 10or less;

R₁ is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)qS(O)_(t)R₄; hydroxy;hydroxy C₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; aryl C₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heterocyclic, heterocyclic C₁₋₄alkyl;heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅;(CR₈R₈)qC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁;C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁(CR₈R₈)qC(O)OR₁₂;(CR₈R₈)qOC(O)R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)qNHS(O)₂R₁₇,(CR₈R₈)qS(O)₂NR₄R₅; or two R₁ moieties together may form O—(CH₂)_(s)O—or a 5 to 6 membered unsaturated ring;

q is 0, or an integer having a value of 1 to 10;

t is 0, or an integer having a value of 1 or 2;

s is an integer having a value of 1 to 3;

v is an integer having a value of 1 to 4;

R₄ and R₅ are independently hydrogen, optionally substituted C₁₋₄ alkyl,optionally substituted aryl, optionally substituted aryl C₁₋₄alkyl,optionally substituted heteroaryl, optionally substituted heteroarylC₁₋₄alkyl, heterocyclic, heterocyclic C₁₄ alkyl, or R₄ and R₅ togetherwith the nitrogen to which they are attached form a 5 to 7 member ringwhich may optionally comprise an additional heteroatom selected fromoxygen, nitrogen or sulfur;

R₆ and R₇ are independently hydrogen or a C₁₋₄ alkyl group, or R₆ and R₇together with the nitrogen to which they are attached form a 5 to 7member ring which ring may optionally contain an additional heteroatomwhich heteroatom is selected from oxygen, nitrogen or sulfur;

Y is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)qS(O)_(t)R₄; hydroxy;hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; heterocyclic,heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅;(CR₈R₈)qC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁;C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁; C(O)R₁₁;(CR₈R₈)qC(O)OR₁₂; (CR₈R₈)qOC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁,(CR₈R₈)qNHS(O)₂R_(d), (CR₈R₈)qS(O)₂NR₄R₅; or two Y moieties together mayform O—(CH₂)_(s)O— or a 5 to 6 membered unsaturated ring;

n is an integer having a value of 1 to 3;

m is an integer having a value of 1 to 3;

R₈ is hydrogen or C₁₋₄ alkyl;

R₁₀ is C₁₋₄ alkyl C(O)₂R₈;

R₁₁ is hydrogen, C₁₋₄ alkyl, optionally substituted aryl, optionallysubstituted aryl C₁₋₄alkyl, optionally substituted beteroaryl,optionally substituted heteroarylC₁₋₄alkyl, optionally substitutedheterocyclic, or optionally substituted heterocyclicC₁₋₄alkyl;

R₁₂ is hydrogen, C₁₋₁₀ alkyl, optionally substituted aryl or optionallysubstituted arylalkyl;

R₁₃ and R₁₄ are independently hydrogen, optionally substituted C₁₋₄alkyl or one of R₁₃ and R₁₄ may be optionally substituted aryl;

R₁₇ is C₁₋₄alkyl, aryl, arylalkyl, heteroaryl, heteroaryl C₁₋₄alkyl,heterocyclic, or heterocyclicC₁₋₄alkyl, wherein the aryl, heteroaryl andheterocyclic rings may all be optionally substituted;

R_(d) is NR₆R₇, alkyl, arylC1-4alklyl, arylC₂₋₄ alkenyl, heteroaryl,hetroaryl-C₁₋₄alkyl, heteroarylC₂₋₄ alkenyl, heterocyclic,heterocyclicC₁₋₄ alkyl, wherein the aryl, heteoaryl and heterocyclicrings may all be optionally substituted;

provided that

R₁ is not 3-phenyl, or 4-isopropyl, or 5-phenyl or 5-ethylsulfonyl, whenR is OH, v is 1, R₁₃ and R₁₄ are hydrogen, and Y is hydrogen;

or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is to a method of treating achemokine mediated disease, wherein the chemokine is one which binds toan IL-8 α or β receptor and which method comprises administering aneffective amount of a compound of Formula (II) or a pharmaceuticallyacceptable salt thereof, as defined herein.

This invention also relates to a method of inhibiting the binding ofIL-8 to its receptors in a mammal in need thereof which comprisesadministering to said mammal an effective amount of a compound ofFormula (II), as defined herein.

This invention also relates to the novel compounds of Formula (II), or apharmaceutically acceptable salt thereof, as defined herein.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of Formula (I) may also be used in association with theveterinary treatment of mammals, other than humans, in need ofinhibition of IL-8 or other chemokines which bind to the IL-8 α and βreceptors. Chemokine mediated diseases for treatment, therapeutically orprophylactically, in animals include disease states such as those notedherein in the Methods of Treatment section.

In compounds of Formula (I), R is suitably any functional moiety whichprovides an ionizable hydrogen having a pKa of 10 or less, preferablyfrom about 3 to 9, more preferably from about 3 to 7. Such functionalgroups include, but are not limited to, hydroxy, carboxylic acid, thiol,—SR₂—OR₂, —NH—C(O)R_(a), —C(O)NR₆R₇, a substituted sulfonamides of theformula —NHS(O)₂R_(b), —S(O)₂NHR_(c), NHC(X₂)NHR_(b), or a tetrazolyl;wherein X₂ is oxygen or sulfur, preferably oxygen. Preferably, thefunctional group is other than a sulfonic acid, either directly or as asubstituent group on the aryl, heteroaryl, or heterocyclic moiety ring,such as in SR₂ or OR₂. More preferably R is OH, SH, or NHS(O)₂R_(b).Suitably, R₂ is a substituted aryl, heteroaryl, or heterocyclic moietywhich ring has the functional moiety providing the ionizable hydrogenhaving a pKa of 10 or less.

Suitably, R₆ and R₇ are independently hydrogen or a C₁₋₄ alkyl group, orR₆ and R₇ together with the nitrogen to which they are attached form a 5to 7 member ring which ring may optionally contain an additionalheteroatom which heteroatom is selected from oxygen, nitrogen or sulfur.This heteroring may be optionally substituted as defined herein.

Suitably R_(a) is an alkyl, aryl, arylC₁₋₄alkyl, heteroaryl,heteroarylC₁₋₄alkyl, heterocyclic, or a heterocyclic C₁₋₄alkyl moiety,all of which may be optionally substituted, as defined herein below.

Suitably, R_(b) is a NR₆R₇, alkyl, aryl, arylC₁₋₄alkyl, arylC₂₋₄alkenyl,heteroaryl, heteroarylC₁₋₄alkyl, heteroarylC₂₋₄ alkenyl, heterocyclic,or heterocyclic C₁₋₄alkyl, or a heterocyclic C₂₋₄alkenyl moiety,camphor, all of which may be optionally substituted one to three timesindependently by halogen; nitro; halosubstituted C₁₋₄ alkyl, such asCF₃; C₁₋₄ alkyl, such as methyl; C₁₋₄ alkoxy, such as methoxy;NR₉C(O)R_(a); C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl. R_(b) is preferablyan optionally substituted phenyl, benzyl, or styryl. When R_(b) is aheteroaryl ring, it is preferably an optionally substituted thiazole, anoptionally substituted thienyl, or an optionally substituted quinolinylring.

Suitably, R₉ is hydrogen or a C₁₋₄ alkyl. Preferably R₉ is hydrogen.When R₉ is the substituent group NR₉C(O)R_(a), then R_(a) is preferablyan alkyl group, such as methyl.

Suitably R_(c) is hydrogen, alkyl, aryl, arylC₁₋₄alkyl, arylC₁₋₄alkenyl,heteroaryl, heteroarylC₁₋₄alkyl, heteroarylC₁₋₄alkenyl, heterocyclic, orheterocyclic C₁₋₄alkyl, or a heterocyclic C₁₋₄alkenyl moiety, all ofwhich may be optionally substituted one to three times independently byhalogen, nitro, halosubstituted C₁₋₄ alkyl, C₁₋₄ alkyl, C₁₋₄ alkoxy,NR₉C(O)R_(a), C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl, wherein R₉ ishydrogen or a C₁₋₄ alkyl. Preferably, R_(c) is an optionally substitutedphenyl.

When R is an OR₂ or SR₂ moiety it is recognized by one of skill in theart that the aryl ring must, therefore, contain the required ionizablehydrogen. The aryl ring may also be additionally substituted,independently, by one to three groups, which groups may also contain anadditional ionizable group, and which include but are not limited to,halogen, nitro, halosubstituted C₁₋₄ alkyl, C₁₋₄ alkyl, C₁₋₄ alkoxy,hydroxy, SH, —C(O)NR₆R₇, —NH—C(O)R_(a), —NHS(O)₂R_(b), S(O)₂NR₆R₇,C(O)OR₈, or a tetrazolyl ring.

In compounds of Formula (I), suitably R₁ is independently selected fromhydrogen; halogen; nitro; cyano; halosubstituted C₁₋₁₀ alkyl, such asCF₃; C₁₋₁₀ alkyl, such as methyl, ethyl, isopropyl, or n-propyl; C₂₋₁₀alkenyl; C₁₋₁₀ alkoxy, such as methoxy, or ethoxy; halosubstituted C₁₋₁₀alkoxy, such as trifluoromethoxy; azide; (CR₈R₈)qS(O)_(t)R₄, wherein tis 0, 1 or 2; hydroxy; hydroxy C₁₋₄alkyl, such as methanol or ethanol;aryl, such as phenyl or naphthyl; aryl C₁₋₄ alkyl, such as benzyl;aryloxy, such as phenoxy; aryl C₁₋₄ alkyloxy, such as benzyloxy;heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀alkenyl; heteroaryl C₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl;(CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₅;(CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁; C₂₋₁₀ alkenylC(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁; C(O)R₁₁; (CR₈R₈)qC(O)OR₁₂;(CR₈R₈)qOC(O)R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)qNHS(O)₂R₁₇,(CR₈R₈)qS(O)₂NR₄R₅; or two R₁ moieties together may form O—(CH₂)_(s)O—or a 5 to 6 membered unsaturated ring; and s is an integer having avalue of 1 to 3. The aryl, arylalkyl, arylalkenyl, heteroaryl,heteroarylalkyl, heteroarylalkenyl, heterocyclic, heterocyclicalkyl, andheterocyclicalkenyl moieties may all be optionally substituted asdefined herein below.

Suitably, q is 0, or an integer having a value of 1 to 10.

When R₁ forms a dioxybridge, s is preferably 1. When R₁ forms anadditional unsaturated ring, it is preferably 6 membered resulting in anaphthylene ring system. This naphthylene ring may be substitutedindependently, 1 to 3 times by the other R₁ moieties as defined above.

Suitably, R₄ and R₅ are independently hydrogen, optionally substitutedC₁₋₄ alkyl, optionally substituted aryl, optionally substituted arylC₁₋₄alkyl, optionally substituted heteroaryl, optionally substitutedheteroaryl C₁₋₄alkyl, heterocyclic, heterocyclicC₁₋₄ alkyl, or R₄ and R₅together with the nitrogen to which they are attached form a 5 to 7member ring which may optionally comprise an additional heteroatomselected from O/N/S.

R₈ is suitably independently selected from hydrogen or C₁₋₄ alkyl.

R₁₀ is suitably C₁₋₁₀ alkyl C(O)₂R₈, such as CH₂C(O)₂H or CH₂C(O)₂CH₃.

R₁₁ is suitably hydrogen, C₁₋₄ alkyl, aryl, aryl C₁₋₄ alkyl, heteroaryl,heteroaryl C₁₋₄alkyl, heterocyclic, or heterocyclic C₁₋₄alkyl.

R₁₂ is suitably hydrogen, C1-10 alkyl, optionally substituted aryl oroptionally substituted arylalkyl.

R₁₇ is suitably C₁₋₄alkyl, aryl, arylalk, heteroaryl,heteroarylC₁₋₄alkyl, heterocyclic, or heterocyclicC₁₋₄alkyl, wherein thearyl, heteroaryl and heterocyclic rings may all be optionallysubstituted.

Preferably R₁ is halogen, cyano, nitro, CF₃, C(O)NR₄R₅, alkenylC(O)NR₄R₅, C(O)R₄R₁₀, alkenyl C(O)OR₁₂, heteroaryl, heteroarylalkyl,heteroaryl alkenyl, or S(O)NR₄R₅, and preferably R₄ and R₅ are bothhydrogen or one is phenyl. A preferred ring substitution for R₁ is inthe 4-position of the phenyl ring.

When R is OH, SH or NHS(O)₂R_(b) then R₁ is preferably substituted inthe 3-position, the 4-position or di substituted in the 3,4-position.The substituent group is suitably an electron withdrawing moiety.Preferably when R is OH, SH or NSO₂R_(b), than R₁ is nitro, halogen,cyano, trifluoromethyl group, C(O)NR₄R₅.

When R is carboxylic acid, then R₁ is preferably hydrogen, or R₁ ispreferably substituted in the 4-position, more preferably substituted bytrifluoromethyl or chloro.

In compounds of Formula (I), suitably R₁₃ and R₁₄ are independentlyhydrogen, optionally substituted C₁₋₄ alkyl which may be straight orbranched as defined herein, or one of R₁₃ and R₁₄ are an optionallysubstituted aryl; v is an integer having a value of 1 to 4.

When R₁₃ or R₁₄ are an optionally substituted alkyl, the alkyl moietymay be substituted one to three times independently by halogen;halosubstituted C₁₋₄ alkyl such as trifluromethyl; hydroxy; hydroxyC₁₋₄alkyl, C₁₋₄ alkoxy; such as methoxy, or ethoxy, halosubstitutedC₁₋₁₀ alkoxy, S(O)_(t)R₄; aryl; NR₄R₅; NHC(O)R₄; C(O)NR₄R₅; or C(O)OR₈.

Suitably, Y is independently selected from hydrogen; halogen; nitro;cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)qS(O)_(t)R₄;hydroxy; hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy;heterocyclic, heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroarylC₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenylC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈;(CR₈R₈)qC(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁;(CR₈R₈)qC(O)OR₁₂; (CR₈R₈)qOC(O)R₁₁; (CR₈R₈)qNR₄C(O)R₁₁,(CR₈R₈)qNHS(O)₂R_(d), (CR₈R₈)qS(O)₂NR₄R₅ or two Y moieties together mayform O—(CH₂)_(s)O or a 5 to 6 membered unsaturated ring When Y forms adioxybridge, s is preferably 1. When Y forms an additional unsaturatedring, it is preferably 6 membered resulting in a naphthylene ringsystem. This naphthylene ring may be substituted 1 to 3 times by other Ymoieties as defined above. The aryl, arylalkyl, arylalkenyl, heteroaryl,heteroarylalkyl, heteroarylalkenyl, heterocyclic, heterocyclicalkyl, andheterocyclicalkenyl moieties noted above may all be optionallysubstituted as defined herein.

Suitably, R_(d) is a NR₆R₇, alkyl, aryl C₁₋₄ alkyl, arylC₂₋₄ alkenyl,heteroaryl, hetroaryl-C₁₋₄alkyl, heteroarylC₂₋₄ alkenyl, heterocyclic,heterocyclicC₁₋₄ alkyl, or heterocyclic C₂₋₄ alkenyl moiety, wherein thearyl, arylalkyl, arylalkenyl, heteroaryl, heteroarylalkyl,heteroarylalkenyl, heterocyclic, and heterocyclicalkyl, andheterocyclicalkenyl moieties noted above may all be optionallysubstituted as defined herein.

Y is preferably a halogen, C₁₋₄ alkoxy, optionally substituted aryl,optionally substituted aryloxy or arylalkoxy, methylene dioxy, NR₄R₅,thio C₁₋₄alkyl, thioaryl, halosubstituted alkoxy, optionally substitutedC₁₋₄ alkyl, or hydroxy alkyl. Y is more preferably mono-substitutedhalogen, disubstituted halogen, mono-substituted alkoxy, disubstitutedalkoxy, methylenedioxy, aryl, or alkyl, more preferably these groups aremono or di-substituted in the 2′-position or 2′-, 3′-position.

While Y may be substituted in any of the 5 ring positions, preferablywhen R is OH, SH, or NHSO₂R_(b), Y is preferably mono-substituted in the2′-position or 3′-position, with the 4′- preferably being unsubstituted.If the ring is disubstituted, when R is OH, SH, or NSO₂R_(b),substituents are preferably in the 2′ or 3′ position of a monocyclicring. While both R₁ and Y can both be hydrogen, it is preferred that atleast one of the rings be substituted, and more preferably that bothrings are substituted.

In compounds of Formula (I), X is suitably oxygen or sulfur, preferablyoxygen.

Exemplified compounds of Formula (I) include:

N-(2-Hydroxy-4-nitrophenyl)-N′-(benzyl)urea

(R)-N-(2-Hydroxy-4-nitrophenyl)-N′-(methylbenzyl)urea

(S)-N-(2-Hydroxy-4-nitrophenyl)-N′-(methylbenzyl)urea

N-[2-hydroxy-3,4-dichlorophenyl]-N′-[benzyl]urea

N-[2-hydroxy-3-cyanophenyl]-N′-[benzyl]urea

N-[2-Hydroxy-5-nitrophenyl]-N′-[benzyl]urea

N-[2-Hydroxy-3-nitrophenyl]-N′-[benzyl]urea

N-[2-benzenesulfonylamino-4-cyanophenyl]-N′-(benzyl)urea

N-Phenethyl-N′-(2-hydroxy-4-nitrophenyl)urea

N-(Diphenylmethyl)-N′-(2-hydroxy-4-nitrophenyl)urea

N-[2-hydroxy-3-nitrophenyl]-N′-[2-chlorobenzyl]urea

N-[1-trifluoromethyl-2-phenyl]N-[2-hydroxy-3-nitrophenyl]urea

N-(2-Hydroxy-4-nitrophenyl)-N′-(2-chlorobenzyl)urea

N-(2-Hydroxy-4-cyanophenyl)-N′-(2-chlorobenzyl)ureaN-(2-Hydroxy-3-trifioromethylphenyl)N′-benzylureaN-[2-hydroxy-3-cyano-4-chlorophenyl]-N′-[benzyl]urea.

As used herein, “optionally substituted” unless specifically definedshall mean such groups as halogen, such as fluorine, chlorine, bromineor iodine; hydroxy; hydroxy substituted C₁₋₁₀alkyl; C₁₋₁₀ alkoxy, suchas methoxy or ethoxy; S(O)_(m′)C₁₋₁₀ alkyl, wherein m′ is 0, 1 or 2,such as methyl thio, methyl sulfinyl or methyl sulfonyl; amino, mono &di-substituted amino, such as in the NR₄R₅ group; NHC(O)R₄; C(O)NR₄R₅;C(O)OH; S(O)₂NR₄R₅; NHS(O)₂R₁₅, C₁₋₁₀ alkyl, such as methyl, ethyl,propyl, isopropyl, or t-butyl; halosubstituted C₁₋₁₀ alkyl, such CF₃; anoptionally substituted aryl, such as phenyl, or an optionallysubstituted arylalkyl, such as benzyl or phenethyl, optionallysubstituted heterocylic, optionally substituted heterocylicalkyl,optionally substituted heteroaryl, optionally substituted heteroarylalkyl, wherein these aryl , hetroaryl, or heterocyclic moieties may besubstituted one to two times by halogen; hydroxy; hydroxy substitutedalkyl; C₁₋₁₀ alkoxy; S(O)_(m′)C₁₋₁₀ alkyl; amino, mono & di-substitutedamino, such as in the NR₄R₅ group; C₁₋₁₀ alkyl, or halosubstituted C₁₋₁₀alkyl, such as CF₃.

R₁₅ is suitably C₁₋₄ alkyl, aryl, aryl C₁₋₄alkyl, heteroaryl,heteroarylC₁₋₄alkyl, heterocyclic, or heterocyclicC₁₋₄alkyl.

Another aspect of the present invention are the novel compounds ofFormula (II), or a pharmaceutically acceptable salt thereof, asdescribed below, which are also useful in inhibiting the binding of IL-8to its receptors in a mammal in need thereof. This invention alsorelates to the pharmaceutical compositions comprising a compound ofFormula (II) and a pharmaceutically acceptable diluent or carrier.Compounds of Formula (II) are also useful for treating a chemokinemediated disease, wherein the chemokine is one which binds to an IL-8 αor β receptor and which method comprises administering an effectiveamount of a compound of Formula (II) or a pharmaceutically acceptablesalt thereof.

Compounds of Formula (II) are represented by the structure:

wherein

X is oxygen or sulfur;

R is any functional moiety having an ionizable hydrogen and a pKa of 10or less;

R₁ is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)qS(O)_(t)R₄; hydroxy;hydroxy C₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; aryl C₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heterocyclic, heterocyclic C₁₋₄alkyl;heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅;(CR₈R₈)qC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁;C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁(CR₈R₈)qC(O)OR₁₂;(CR₈R₈)qOC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)qNHS(O)₂R₁₇,(CR₈R₈)qS(O)₂NR₄R₅; or two R₁ moieties together may form O—(CH₂)_(s)O—or a 5 to 6 membered unsaturated ring;

q is 0, or an integer having a value of 1 to 10;

t is 0, or an integer having a value of 1 or 2;

s is an integer having a value of 1 to 3;

v is an integer having a value of 1 to 4;

R₄ and R₅ are independently hydrogen, optionally substituted C₁₋₄ alkyl,optionally substituted aryl, optionally substituted aryl C₁₋₄alkyl,optionally substituted heteroaryl, optionally substituted heteroarylC₁₋₄alkyl, heterocyclic, heterocyclic C₁₋₄ alkyl, or R₄ and R₅ togetherwith the nitrogen to which they are attached form a 5 to 7 member ringwhich may optionally comprise an additional heteroatom selected fromO/N/S;

R₆ and R₇ are independently hydrogen or a C₁₋₄ alkyl group, or R₆ and R₇together with the nitrogen to which they are attached form a 5 to 7member ring which ring may optionally contain an additional heteroatomwhich heteroatom is selected from oxygen, nitrogen or sulfur;

Y is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)qS(O)_(t)R₄; hydroxy;hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; heterocyclic,heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅;(CR₈R₈)qC(O)NR₄R₅; (CR₈R₈)qC(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁;C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁; C(O)R₁₁;(CR₈R₈)qC(O)OR₁₂; (CR₈R₈)qOC(O)R₁₁; (CR₈R₈)qNR₄C(O)R₁₁,(CR₈R₈)qNHS(O)₂R_(d), (CR₈R₈)qS(O)₂NR₄R₅; or two Y moieties together mayform O—(CH₂)_(s)O— or a 5 to 6 membered unsaturated ring;

n is an integer having a value of 1 to 3;

m is an integer having a value of 1 to 3;

R₈ is independently selected from hydrogen or C₁₋₄ alkyl;

R₁₀ is C₁₋₁₀ alkyl C(O)₂R₈;

R₁₁ is hydrogen, C₁₋₄ alkyl, optionally substituted amyl, optionallysubstituted aryl C₁₋₄alkyl, optionally substituted heteroaryl,optionally substituted heteroarylC₁₋₄alkyl, optionally substitutedheterocyclic, or optionally substituted heterocyclicC₁₋₄alkyl;

R₁₂ is hydrogen, C₁₋₁₀ alkyl, optionally substituted aryl or optionallysubstituted arylalkyl;

R₁₃ and R₁₄ are independently hydrogen, optionally substituted C₁₋₄alkyl, or one of R₁₃ and R₁₄ may be optionally substituted aryl;

R₁₇ is C₁₋₄alkyl, aryl, arylalkyl, heteroaryl, heteroarylC₁₋₄alkyl,heterocyclic, or heterocyclicC₁₋₄alkyl, wherein the aryl, heteroaryl andheterocyclic rings may all be optionally substituted;

R_(d) is NR₆R₇, alkyl, arylC₁₋₄alklyl, arylC₂₋₄ alkenyl, heteroaryl,hetroaryl-C₁₋₄alkyl, heteroarylC₂₋₄ alkenyl, heterocyclic,heterocyclicC₁₋₄ alkyl, wherein the aryl, heteoaryl and heterocyclicrings may all be optionally substituted;

E is optionally selected from

 the asterix * denoting point of attachment of the ring; and

Z is optionally selected from

 the asterix * denoting point of attachment of the ring, with at leastone E ring or Z ring being present;

or a pharmaceutically acceptably salt thereof.

Suitably, the variables for Formula (II), such as X, R, R₁, R₂, R₃, R₄,R₅, R₆, R₇, R₈, R₉, Y, R_(a), R_(b), R_(c), n, m, s, t, and v terms,etc. are as defined in Formula (I) above. The E ring and Z ring aredenoted by its point of attachment through the asterix (*) mayoptionally be present. If it is not present the ring is a phenyl moietywhich is substituted by the R and R₁ terms as shown. At least one E or Zring is necessary. The E ring may be substituted by the R₁ moiety in anyring, saturated or unsaturated, and is shown for purposes hereinsubstituted only in the unsaturated ring(s). The Z ring may besubstituted by the Y moiety in any ring, saturated or unsaturated, andis shown for purposes herein substituted only in the unsaturatedring(s).

Suitable compounds of Formula (II) include:

N-(Hydroxy-3-naphthyl)-N′-(benzyl)urea

Suitable pharmaceutically acceptable salts are well known to thoseskilled in the art and include basic salts of inorganic and organicacids, such as hydrochloric acid, hydrobromic acid, sulphuric acid,phosphoric acid, methane sulphonic acid, ethane sulphonic acid, aceticacid, malic acid, tartaric acid, citric acid, lactic acid, oxalic acid,succinic acid, fumaric acid, maleic acid, benzoic acid, salicylic acid,phenylacetic acid and mandelic acid In addition, pharmaceuticallyacceptable salts of compounds of Formula (I) may also be formed with apharmaceutically acceptable cation, for instance, if a substituent groupcomprises a carboxy moiety. Suitable pharmaceutically acceptable cationsare well known to those skilled in the art and include alkaline,alkaline earth, ammonium and quaternary ammonium cations.

The following terms, as used herein, refer to:

“halo”—all halogens, that is chloro, fluoro, bromo and iodo.

“C₁₋₁₀alkyl” or “alkyl”—both straight and branched chain radicals of 1to 10 carbon atoms, unless the chain length is otherwise limited,including, but not limited to, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl and the like.

The term “cycloalkyl” is used herein to mean cyclic radicals, preferablyof 3 to 8 carbons, including but not limited to cyclopropyl,cyclopentyl, cyclohexyl, and the like.

The term “alkenyl” is used herein at all occurrences to mean straight orbranched chain radical of 2-10 carbon atoms, unless the chain length islimited thereto, including, but not limited to ethenyl, 1-propenyl,2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl and the like.

“aryl”—phenyl and naphthyl;

“heteroaryl” (on its own or in any combination, such as “heteroaryloxy”,or “heteroaryl alkyl”)—a 5-10 membered aromatic ring system in which oneor more rings contain one or more heteroatoms selected from the groupconsisting of N, O or S, such as, but not limited, to pyrrole, pyrazole,furan, thiophene, quinoline, isoquinoline, quinazolinyl, pyridine,pyrimidine, oxazole, thiazole, thiadiazole, triazole, imidazole, orbenzimidazole.

“heterocyclic” (on its own or in any combination, such as“heterocyclicalkyl”)—a saturated or partially unsaturated 4-10 memberedring system in which one or more rings contain one or more heteroatomsselected from the group consisting of N, O, or S; such as, but notlimited to, pyrrolidine, piperidine, piperazine, morpholine,tetrahydropyran, or imidazolidine.

The term “arylalkyl” or “heteroarylalkyl” or “heterocyclicalkyl” is usedherein to mean C₁₋₁₀ alkyl, as defined above, attached to an aryl,heteroaryl or heterocyclic moiety, as also defined herein, unlessotherwise indicated.

“sulfinyl”—the oxide S(O) of the corresponding sulfide, the term “thio”refers to the sulfide, and the term “sulfonyl” refers to the fullyoxidized S(O)₂ moiety.

The term “wherein two R₁ moieties (or two Y moieties) may together forma 5 or 6 membered unsaturated ring” is used herein to mean the formationof a napthylene ring system or a phenyl moiety having attached a 6membered partially unsaturated ring such as a C₆ cycloalkenyl, i.ehexene, or a C₅ cyloalkenyl moiety, cyclopentene.

The compounds of Formula (I), and (II) may be obtained by applyingsynthetic procedures, some of which are illustrated in the Schemesbelow. The synthesis provided for in these Schemes is applicable for theproducing compounds of Formula (I),and (II) having a variety ofdifferent R, R₁, and Aryl groups which are reacted, employing optionalsubstituents which are suitably protected, to achieve compatibility withthe reactions outlined herein. Subsequent deprotection, in those cases,then affords compounds of the nature generally disclosed. Once the ureanucleus has been established, further compounds of these formulas may beprepared by applying standard techniques for functional groupinterconversion, well known in the art. While the schemes are shown withcompounds only of Formula (I) this is merely for illustration purposesonly.

Ortho substituted phenyl ureas shown in 2-scheme 1 may be prepared bystandard conditions involving the condensation of commercially availableortho substituted aniline(Aldrich Chemical Co., Milwaukee, Wis.) withthe commercially available optionally substituted aryl isocyanate(Aldrich Chemical Co., Milwaukee, Wis.) in an aprotic solvent (DMF,toluene). When the 1-(RSO₂NH)2-(NH₂)Ph is not commercially available itcan be made by treating the commercially available RSO₂Cl with thecorresponding 2-phenylene diamine in the presence of an base liketriethyl amine or NaH in an aprotic solvent (like methylene chloride orDMF).

If the desired 2-substituted aniline 5-scheme 2, is not commerciallyavailable the corresponding nitro compound can be prepared from 3-scheme2, under standard nitration conditions (using HNO₃ or BF₄NO₃) at 23° C.The nitro compound is then reduced to the corresponding aniline usingSnCl₂ in EtOH (or alternately H₂/Pd or LiAlH₄).

If the desired 2-amino benzenethiol 8-scheme 3 is not commerciallyavailable it can be synthesized by reaction of the phenyl aniline withthe thiocyanate anion in the presence of an oxidant(like bromine) toproduce the 2-amino benzthiazole 7-scheme 3. This thiazole can then behydrolyzed to the desired 2-amino benzenethiol 8-scheme 4 with a strongbase like NaOH in a protic solvent (i.e., EtOH).

In the case where the thioisocyanate or phenyl isocyanate is notcommercially available, the thiourea or urea 11-scheme 4 may be preparedfrom the commercially available ortho substituted aniline. This compoundis first protected with a protecting group (tert-butyl dimethyl silyl orbenzyl) by conditions well known in the art (see Greene, T ProtectingGroups in Organic Synthesis Wiley&Sons, New York, 1981). This protectedaniline is then reacted, in the presence of a base(like triethyl amineor sodium bicarbonate), with either thiophosgene or a solution ofphosgene in an aprotic solvent (i.e. DMF, toluene), followed by anilineto produce the protected thiourea or urea respectively. Thecorresponding urea or thiourea is then deprotected, using conditionsstandard in the art, to form the desired thiourea or urea 11-scheme 4.

Pharmaceutically acceptable salts of compounds of Formula (I) may beobtained in known manner, for example by treatment thereof with anappropriate amount of acid or base in the presence of a suitablesolvent.

Numerous conversions of aryl halides to aryl cyano derivatives withcopper (I) cyanide have been published. However, no examples of an arylring with a hydroxy group present were mentioned. Several attempts toobtain a cyano phenol moiety with published results failed. Using knownconditions of elevated temperatures, greater than 170° C., such as from180 to 210° did not yield displacment of the halogen to a cyano moiety.Standard bases, such as DMF and pyridine further provided no desiredproduct. Intermediates such as 2-amino-5-fluorophenol,2-nitro5-fluorophenol, 2-nitro-5-methyl6-bromophenol were tried with achange of halogens, from fluorine to chlorine to bromine, and with useof copper (I) cyanide. The use of a bromine derivative, such as2-nitro-5-methyl-6-bromophenol, with dimethylformamide and usingtriethylamine with a catalytic amount of dimethylamino pyridine andcopper (I) cyanide at reduced temperatures, i.e. <100° C., preferably 60to about 80° C. for reduced times from strandarized procedures, i.e.,<18 hours, preferably about 4 to 6 hours yielded the desired products.

In the Examples, all temperatures are in degrees Centigrade (° C.). Massspectra were performed upon a VG Zab mass spectrometer using fast atombombardment, unless otherwise indicated. ¹H-NMR (hereinafter “NMR”)spectra were recorded at 250 MHz or 400 MHz using a Bruker AM 250 or Am400 spectrometer, respectively. Multiplicities indicated are: s=singlet,d=doublet, t=triplet, q=quartet, m=multiplet and br indicates a broadsignal. Sat. indicates a saturated solution, equiv. indicates theproportion of a molar equivalent of reagent relative to the principalreactant.

Flash chromatography is run over Merck Silica gel 60 (230-400 mesh).

SYNTHETIC EXAMPLES

The invention will now be described by reference to the followingexamples which are merely illustrative and are not to be construed as alimitation of the scope of the present invention. All temperatures aregiven in degrees centigrade, all solvents used herein are of the highestavailable purity and all reactions are run under anhydrous conditions inan argon atmosphere unless otherwise indicated.

General Method A: Synthesis of N-phenyl, N′-phenyl urea

To a solution of phenyl isocyanate (1.0 equiv.) in dimethyl formamide (1milliliters (hereinafter “ml”))) the corresponding aniline (1.0 equiv.)was added. The reaction mixture was stirred at 80° C. until complete(24-48 hours (hereinafter “hrs” or “h”)), then the solvent was removedunder vacuum. The purifications, yields and spectral characteristics foreach individual compound are listed below.

General Method B:Synthesis of sulfonamide

The ortho substituted aniline (1 equiv.), triethyl amine (1 equiv.) andthe desired sulfonyl chloride (1 equiv.) were combined in methylenechloride and allowed to stir at about 23° C. until complete (12-36h).The reaction mixture was partitioned between water and methylenechloride. The organic layer was separated and dried over magnesiumsulfate, filtered and concentrated in vacuo. The purifications of eachcompound are listed below.

EXAMPLE 1 Preparation of N-(2-hydroxy-4-nitrophenyl)-N′-(benzyl)urea

N-(2-Hydroxy-4-nitrophenyl)-N′-(benzyl)urea was prepared from 2-hydroxy4-nitro aniline (308 mg, 2.0 mmol) and benzyl isocyanate (2 mmol)according to the procedure in General Method A. The product was purifiedby dilution with methylene chloride and precipitation with hexanes.Filtering afforded the title compound (362 mg, 52%). EI-MS m/z 288(M+H)⁺

EXAMPLE 2 Preparation of (R)-N-(2-hydroxy-4-nitrophenyl)-N′-(methylbenzyl)urea

(R)-N-(2-Hydroxy4-nitrophenyl)-N′-(methylbenzyl)urea was prepared from2-hydroxy 4-nitro aniline (308 mg, 2.0 mmol) and (R)-methyl benzylisocyanate (2 mmol) according to the procedure in General Method A. Theproduct was purified by dilution with methylene chloride andprecipitation with hexanes. Filtering afforded the title compound (427mg, 71%). EI-MS m/z 300(M−H)⁻

EXAMPLE 3 Preparation of (S)-N-(2-hydroxy-4-nitrophenyl)-N′-(methylbenzyl)urea

(S)-N-(2-Hydroxy-4-nitrophenyl)-N′-(methylbenzyl)urea was prepared from2-hydroxy 4-nitro aniline (308 mg, 2.0 mmol) and (S)-methyl benzylisocyanate (2 mmol) according to the procedure in General Method A. Theproduct was purified by dilution with methylene chloride andprecipitation with hexanes. Filtering afforded the title compound (462mg, 76%). EI-MS m/z 300(M−H)⁻

EXAMPLE 4 Preparation of N-[2-hydroxy-3,4 -dichlorophenyl]-N′-[benzyl]urea a)Preparation of 2-nitro-5,6 dichlorophenol

2,3 dichlorophenol (3.26 g, 20 mmol) was dissolved in methylenechloride(40 ml) followed by the addition of sodium nitrate (1.88 g, 22mmol). The addition of sulfuric acid (20 ml/3M) was then made, followedby addition of a catalytic amount of sodium nitrite. The mixture wasallowed to stir. After 24 hrs, the reaction mixture was diluted withmethylene chloride and extracted with water. The organic layer was driedover MgSO₄ and filtered. The solvent was evaporated and chromatographyof the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gave the desiredproduct(1.8 g, 44%). ¹H NMR (CD₃COCD₃): δ8.04 (d,1H), 7.15 (d, 1H).

b)Preparation of 2-amino-5,6 dichlorophenol

A mixture of 5,6-dichloro-2-nitrophenol(1.8 g, 8.7 mmol) and tin (II)chloride (5.8 g, 26.1 mmol) in ethanol(50 ml) was heated at 80° C. underargon. After 2 hours, the starting material had disappeared and thesolution was allowed to cool down and then poured into ice. The pH wasmade slightly basic (pH7-8), by addition of solid NaOH, before beingextracted with ethyl acetate. The organic phase was washed with brine,dried over MgSO₄ and filtered. The solvent was evaporated andchromatography of the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gavethe desired product(1.4 mg, 90%). ¹H NMR (CD₃OD): δ6.71 (d, 1H), 6.45(d, 1H).

c)Preparation of N-[2-hydroxy-3,4-dichlorophenyl]-N′-[benzyl] urea

N-[2-hydroxy-3,4-dichlorophenyl]-N′-[benzyl] urea was prepared from2-amino-5,6dichlorophenol (80 mg, 0.50 mmol) according to the procedurein General Method A. The product was purified by precipitation frommethylene chloride/hexane(1 equiv./20 equiv.) and filtering. (120 mg,77%) ¹H NMR (CD₃OD): δ7.72 (d, 1H), 7.40-7.25 (m, 5H), 6.97 (d, 1H),4.40 (s, (2H).

EXAMPLE 5 Preparation of N-[2-hydroxy-3-cyanophenyl]-N′-[benzyl] ureaa)Preparation of 2-nitro-6cyanophenol

2-Cyanophenol (4.72 g, 40 mmol) was dissolved in methylene chloride(80ml) followed by the addition of sodium nitrate (3.76 g, 44 mmol). Theaddition of sulfuric acid (40 ml/ 3M) was then made, followed byaddition of a catalytic amount of sodium nitrite. The mixture wasallowed to stir. After 24 hrs, the reaction mixture was diluted withmethylene chloride and extracted with water. The organic layer was driedover MgSO₄ and filtered. The solvent was evaporated and chromatographyof the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gave the desiredproduct(2.8 g, 42%). ¹H NMR (CD₃COCD₃): δ8.47 (d, 1H), 8.15 (d, 1H),7.30 (t, 1H).

b)Preparation of 2-amino-5-cyanophenol

A mixture of 6cyano2-nitrophenol(2.40 g, 14.6 mmol) and tin (II)chloride (9.6 g, 43.2 mmol) in acetic acid(100 mL) was heated at 80° C.under argon. After 2 hours, the starting material had disappeared andthe solution was allowed to cool down and then poured into ice. The pHwas made slightly basic (pH 7-8), by addition of solid NaOH, beforebeing extracted with ethyl acetate. The organic phase was washed withbrine, dried over MgSO₄ and filtered. The solvent was evaporated andchromatography of the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gavethe desired product(1.50 g, 77%). ¹H NMR (CD₃OD): δ6.92 (d, 1H),6.85-6.69 (m,2H).

c)Preparation of N-[2-hydroxy-3-cyanophenyl]-N′-[benzyl] urea

N-[2-hydroxy-3-cyanophenyl]-N′-[benzyl] urea was prepared from2-amino-6-cyanophenol (134 mg, 1.00 mmol) according to the procedure inGeneral Method A. The product was purified by precipitation frommethylene chloride/hexane(1 equiv./20 equiv.) and filtering. (220 mg,82%). ¹H NMR (CD₃OD): δ7.61 (d, 1H), 7.40-7.21 (m, 6H), 6.92 (t, 1H),4.40 (s, 2H). cl EXAMPLE 6

Preparation of N-[2-hydroxy-5-nitrophenyl]-N′[benzyl] urea

N-[2-Hydroxy-5-nitrophenyl]-N′-[benzyl] urea was prepared from2-amino-4-nitrophenol (154 mg, 1.00 mmol) according to the procedure inGeneral Method A. The product was purified by precipitation frommethylene chloride/hexane(1 equiv./20 equiv.) and filtering (240 mg,84%). ¹H NMR (CD₃OD): δ9.03 (s, 1H), 7.82 (d, 1H), 7.50-7.22 (m, 5H),6.94 (d, 1H), 4.45 (s, 2H).

EXAMPLE 7 Preparation of N-[2-hydroxy-3-nitrophenyl]-N′-[benzyl] urea

N-[2-Hydroxy-3-nitrophenyl]-N′-[benzyl] urea was prepared from2-amino-6-nitrophenol (150 mg, 1.0 mmol) according to the procedure inGeneral Method B. The product was purified by precipitation frommethylene chloride/hexane(1 equiv./20 equiv.) and filtering. (235 mg,82%). ¹H NMR (CD₃OD): δ8.39 (d, 1H), 7.40-7.28 (m, 5H), 6.97 (t, 1H),4.40 (s, 2H).

EXAMPLE 8 Preparation ofN-[2-benzenesulfonylamino-4cyanophenyl]-N′-[benzyl] urea a)Synthesis of3-(phenylsulfamido) benzonitrile

The of 3-(phenylsulfamido) benzonitrile was synthesized from the 3-cyanoaniline (23.9 g, 0.2 mol) by Method B. It was purified byrecrystalization from EtOH (15.8 g, 31%).¹H NMR (CDC₃): δ7.95(s, 1H),7.84 (d, 2H, J=8.0 Hz), 7.59 (t, 1H, J=8.0 Hz), 7.45 (m, 2H), 7.35 (m,4H).

b)Synthesis of 3-(phenylsulfamido) 4-nitro benzonitrile

The 3-(phenylsulfamido) benzonitrile(10 g, 39 mmol) was dissolved inacetic anhydride and treated with concentrated nitric acid dropwise atroom temperature until all the starting material had been consumed. Thereaction mixture was quenched by carefully pouring it into sodiumbicarbonate and then left to sit until all gas evolution had subsided.It was partitioned between methylene chloride and water. The organiclayer was dried over sodium sulfate and filtered. The reaction mixturewas concentrated in vacuo, absorbed onto silica gel and purified bycolumn chromatography(methylene chloride/hexane) to afford the titlecompound (1.7 g, 15%). EI-MS m/z 302(M+H)⁺

c)Synthesis of 3-(phenylsulfamido) 4-amino benzonitrile

The 3-(phenylsulfamido) 4nitro benzonitrile(1.5 g, 4.9 mmol) was treatedwith tin chloride dihydrate in EtOH at 80° C. for 12 h. It was thenconcentrated and flushed through a plug of silica gel with 5%methanol/methylene chloride. The filterate was absorbed onto silica geland purified by flash chromatography(ethyl acetate/hexane) to afford thetitle compound (0.9 g, 60%). EI-MS m/z 274 (M+H)⁺

d)Synthesis of N-[2-benzenesulfonylamino4-cyanophenyl]-N′-[2-benzyl]urea

N-[2-benzenesulfonylamino-4-cyanophenyl]-N′-[2-benzyl] urea was preparedfrom 2-benzenesulfonylamino4cyanoaniline (71 mg, 0.26 mmol) according tothe procedure in General Method A. The product was purified byprecipitation from methylene chloride/hexane(1 equiv./2 equiv.) andfiltering. (79 mg, 72%). ¹H NMR (CD₃OD): δ8.15 (d, 1H), 7.70-7.40 (m,6H), 7.35-7.15 (m, 5H),6.75 (s, 1H), 4.37 (s, 2H).

EXAMPLE 9 Preparation of N-Phenylethyl-N′-(2-hydroxy-4-nitrophenyl)urea

To a solution of phenethyl isocyanate (382 mg, 2.6 mmol) in toluene,2-amino-5-nitrophenol (400 mg, 2.6 mmol) was added. The reaction mixturewas stirred at 80° C. for 24 hours, then cooled to room temperature. Theproduct was purified by precipitation from toluene and filtering (650mg, 83%). m.p: 188.6-189.4° C.; EI-MS m/z 302 (M+H)⁺.

EXAMPLE 10 Preparation ofN-(Diphenylmethyl)-N′-(2-hydroxy-4nitrophenyl)urea

To a solution of aminodiphenylmethane (200 mg, 1.09 mmol) in toluene,triethylamine (0.18 ml, 1.11 mmol) and triphosgene (129.4 mg, 0.44 wereadded. The reaction mixture was stirred at 80° C. for 3 hours. It wascooled to room temperature and all solvent was evaporated. Then2-amino5-nitrophenol (168 mg, 1.09 mmol) in DMF (2 mL) was added. Thereaction mixture was stirred at 80° C. for 16 hours, then cooled to roomtemperature. Chromatography of the resulting liquid on silica gel(hexane:ethyl acetate; 1:1) gave product (210 mg, 53%). m.p:237.4-237.9° C.; EI-MS m/z 364 (M+H)⁺.

EXAMPLE 11 Preparation of N-[1-trifluoromethyl-2-phenyl]N-[2-hydroxy-3-nitrophenyl] urea, N-(1-trifluoromethyl- 1-phenylmethane)-N′-(2-hydroxy-4-nitrophenyl)urea a) Preparation ofα-(trifluoromethyl)benzylmesylate

To a solution of α-(trifluoromethyl)benzylalcohol (2.0 g, 11.35 mmol) inCH₂Cl₂ (20 mL), methanesulfonyl chloride (1.3 mL, 17.0 mmol) andtriethylamine (5.8 mL, 40 mmol) were added. The reaction mixture wasstirred at room temperature for 16 hours. All the solvent was evaporatedto give the desired product (2.8 g, 97%). EI-MS m/z 255 (M⁺).

b) Preparation of α-(trifluoromethyl)benzylazide

To a solution of α-(trifluoromethyl)benzylmesylate (2.8 g, 11.0 mmol) inDMF (15 mL), sodium azide (7.38 g, 110 mmol) was added. The reactionmixture was stirred at 60° C. for 48 hours. Then the reaction mixturewas partitioned between ethyl acetate and water. The organic layer wasdried over MgSO₄ and filtered. The solvent was evaporated andchromatography of the resulting solid on silica gel gave the desiredproduct (849 mg, 37%). EI-MS m/z 202 (M⁺).

c) Preparation of α-(trifluoromethyl)benzylamine

To a solution of α-(trifluoromethyl)benzylazide (849 mf, 4.22 mmol) inether (10 mL), lithium aluminum hydride (160 mg, 12.7 mmol) was added.The reaction mixture was stirred at room temerature for 2 hours. Then0.46 ml H₂O, 0.51 mL of 15% NaOH and 1.06 mL of H₂O were added. Thesolid was filtered. The liquid was concentrated under reduced pressureto give desired product (647 mg, 88%). EI-MS m/z 176 (M⁺).

d) Preparation of N-(1-trifluoromethyl-1-phenylmethane)-N′-(2-hydroxy-4-nitrophenyl)urea

To a solution of 2-tert-butyldimethylsilyloxy4-nitroaniline (200 mg,0.75 mmol) in toluene (5 mL), triphosgene (88.4 mg, 0.3 mmol) andtriethylamine (0.13 mL, 0.9 mmol) were added The reaction mixture wasstirred at 80° C. for 3 hours, Then was cooled to room temperature andevaporated all solvent. The residue was dissolved in DMF (1 mL).α-(trifluoromethyl)benzylamine (131 mg, 0.75 mmol) was added. Thereaction mixture was stirred at 80° C. for 16 hours Chromatography ofthe resulting liquid on silica gel gave desired product (150 mg, 57%).EI-MS m/z 356 (M⁺); m.p, 111.5-112.4.

EXAMPLE 12 Preparation ofN-(2-Hydroxy4-nitrophenyl)-N′-(2-chlorobenzyl)urea

To a solution of 2-tert-butyldimethylsilyloxy-4-nitroaniline (300 mg,1.12 mmol) in toluene (10 mL), triphosgene (126 mg, 0.45 mmol) andtriethylamine (0.195 mL, 1.34 mmol) were added. The reaction mixture wasstirred at 80° C. for 3 hours. Then the reaction mixture was cooled toroom temperature and all the solvent was evaporated. The residue wasdissolved in DMF (1 mL). 2-chlorobenzylamine (158.6 mg, 1.12 mmol) wasadded. The reaction mixture was stirred at 80° C. for 16 hoursChromatography of the resulting liquid on silica gel (30% Ethylacetate/Hexane) gave desired product (210 mg, 57%). EI-MS m/z 322.8(M⁺); m.p. 199.2-199.6° C.

EXAMPLE 13 Preparation ofN-(2-Hydroxy-4cyanophenyl)-N′-2chlorobenzyl)urea a) Preparation of2chlorobenzylisocyanate

To a solution of 2-chlorobenzylamine (1.0 g, 7.06 mmol) in a mixture ofCH₂Cl₂ and H₂O (10 mL/10 mL), triphosgene (838 mg, 2.82 mmol) and NaHCO₃(1.78 g, 21.2 mmol) were added. The reaction mixture was stirred at roomtemperature for 16 hours. Then the reaction mixture was partitionedbetween CH₂Cl₂ and water. The combined organic layers are dried overMgSO₄ and filtered. The solvent was evaporated to give the desiredproduct (1.1 g, 93%). EI-MS m/z 168.7 (M⁺).

b) Preparation of N-(2-Hydroxy-4-cyanophenyl)N′-(2-chlorobenzyl)urea

To a solution of 2-chlorobenzylisocyanate (125.6 mg, 0.75 mmol) in DMF(1.0 ml), 2-hydroxy-4-cyanoaniline (100 mg, 0.75 mmol) was added. Thereaction mixture was stirred at 80° C. for 16 hours. Chromatography ofthe resulting liquid on silica gel (50% Ethyl acetate/Hexane) gavedesired product (80 mg, 58.8%). EI-MS m/z 302.8 (M⁺).

EXAMPLE 14 Preparation ofN-(2-Hydroxy-3-trifluoromethylphenyl)-N′-benzylurea a) Preparation of3-trifluoromethyl-2-hydroxyaniline

To the solution of 2-trifluoromethyl-6-nitrophenol (4.2 g, 20.3 mmol) inethanol (20 ml, Tin (II) chloride (22.9 g, 101 mmol) was added. Thereaction mixture was stirred at reflux for about 4 hours, tThen it wascooled to room temperature. The NaHCO₃ (aq) was added until pH=7 wasobtained and then the mixture was extracted with ethyl acetate (3×). Thecombined organic layer was dried over MgSO₄, filtered and concentratedunder reduced pressure to give desired product (3.4 g, 94%). EI-MS m/z178 (M⁺).

b) Preparation of N-(2-Hydroxy-3-trifluoromethylphenyl)-N′-benzylurea

To a solution of benzylisocyanate (74.56 mg, 0.56 mmol) in DMF (1.0 ml),2-hydroxy-3-trifluoromethylaniline (100 mg, 0.56 mmol) was added. Thereaction mixture was stirred at 80° C. for 16 hours. Chromatography ofthe resulting liquid on silica gel gave desired product (140 mg, 80%).EI-MS m/z 311.1 (M⁺).

EXAMPLE 15 Preparation ofN-[2-hydroxy-3-cyano-4-chlorophenyl]-N′-[benzyl] urea a) Preparation of2-cyano-3-chlorophenol

2-Benzoyl-6-chlorobenzonitrile (10.0 g, 41 mmol) was dissolved inmethylene chloride(40 ml) and trifluoroacetic acid(40 ml) and stirred atroom temperature for 24 hrs. The reaction mixture is diluted withmethylene chloride and extracted with water. The organic layer is driedover MgSO₄ and filtered. The solvent was evaporated and chromatographyof the resulting solid on silica gel (4%MeOH/CHCl₂) gave the desiredproduct(4.2 g, 67%). ¹H NMR (CD₃COCD₃): δ10.31 (s,1H), 7.51 (t, 1H),7.11 (d, 1H) 7.05 (d, 1H).

b) Preparation of 2-nitro-5-chloro-6cyanophenol

2-cyano-3-chlorophenol(2.8 g, 18.3 mmol) was dissolved in methylenechloride(40 ml) followed by the addition of sodium nitrate (1.70 g, 20.1mmol). The addition of sulfuric acid (15 ml/ 3M) is then made, followedby addition of a catalytic amount of sodium nitrite. The mixture isallowed to stir. After 24 hrs, the reaction mixture is diluted withmethylene chloride and extracted with water. The organic layer is driedover MgSO₄ and filtered. The solvent was evaporated and chromatographyof the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gave the desiredproduct(1.05 g, 29.2%). ¹H NMR (CD₃COCD₃): δ8.09 (d,1H), 6.55 (d, 1H).

c) Preparation of 2-amino-5-chloro6-cyanophenol

A mixture of 2-nitro-5-chloro-6-cyanophenol (750 mg, 3.78 mmol) and tin(II) chloride (2.6 g, 11.4 mmol) in ethanol(50ml) is heated at 80° C.under argon. After 2 hours, the starting material has disappeared andthe solution is allowed to cool down and then poured into ice. The pH ismade slightly basic (pH7-8), by addition of solid NaOH, before beingextracted with ethyl acetate. The organic phase was washed with brine,dried over MgSO₄ and filtered. The solvent was evaporated andchromatography of the resulting solid on silica gel (4%MeOH/CH₂Cl₂) gavethe desired product(410 mg, 65%). ¹H NMR (CD₃0D): δ8 6.89 (d, 1H), 6.88(d,1H).

d)Preparation of N-[2-hydroxy-3cyano-4-chlorophenyl]-N′-[benzyl] urea

N-[2-hydroxy-3-cyanophenyl]-N′-[benzyl] urea was prepared from2-amino-5-chloro-6-cyanophenol (50 mg, 0.30 mmol) according to theprocedure in General Method B. The product was purified by precipitationfrom methylene chloride/hexane(1 equiv./20 equiv.) and filtering. (69mg, 77%). ¹H NMR (CD₃SO₂CD₃): δ11.94 (s, 1H), 8.64 (s, 1H), 7.89 (d,1H), 7.41 (t, 1H), 7.34 (s, 1H), 7.25-7.18 (m, 5H), 7.04 (d, (1H).

METHOD OF TREATMENT

The compounds of Formula (I) and (II), or a pharmaceutically acceptablesalt thereof can be used in the manufacture of a medicament for theprophylactic or therapeutic treatment of any disease state in a human,or other mammal, which is exacerbated or caused by excessive orunregulated IL-8 cytokine production by such mammal's cell, such as butnot limited to monocytes and/or macrophages, or other chemokines whichbind to the IL-8 α or β receptor, also referred to as the type I or typeII receptor.

Accordingly, the present invention provides a method of treating achemokine mediated disease, wherein the chemokine is one which binds toan IL-8 α or β receptor and which method comprises administering aneffective amount of a compound of Formula (I) or (II) or apharmaceutically acceptable salt thereof. In particular, the chemokinesare IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78.

For purposes herein, the compounds of Formula (I) and (II) all have thesame dosages, and formulations as that of Formula (I) are usedinterchangeably.

The compounds of Formula (I) are administered in an amount sufficient toinhibit cytokine function, in particular IL-8, GROα, GROβ, GROγ, NAP-2or ENA-78, such that they are biologically regulated down to normallevels of physiological function, or in some case to subnormal levels,so as to ameliorate the disease state. Abnormal levels of IL-8, GROα,GROβ, GROγY, NAP-2 or ENA-78 for instance in the context of the presentinvention, constitute: (i) levels of free IL-8 greater than or equal to1 picogram per mL; (ii) any cell associated IL-8, GROα, GROβ, GROγ,NAP-2 or ENA-78 above normal physiological levels; or (iii) the presenceIL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 above basal levels in cells ortissues in IL-8, GROα, GROβ, GROγ, NAP-2 or ENA-78 respectively, isproduced.

There are many disease states in which excessive or unregulated IL-8production is implicated in exacerbating and/or causing the disease.Chemokine mediated diseases include psoriasis, atopic dermatitis,arthritis, asthma, chronic obstructive pulmonary disease, adultrespiratory distress syndrome, inflammatory bowel disease, Crohn'sdisease, ulcerative colitis, stroke, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, cardiac and renal reperfusioninjury, glomerulonephritis, thrombosis, graft vs. host reaction,alzheimers disease, allograft rejections, malaria, restinosis,angiogenesis or undesired hematopoietic stem cells release.

These diseases are primarily characterized by massive neutrophilinfiltration, T-cell infiltration, or neovascular growth, and areassociated with increased IL-8, GROα, GROβ, GROγ or NAP-2 productionwhich is responsible for the chemotaxis of neutrophils into theinflammatory site or the directional growth of endothelial cells. Incontrast to other inflammatory cytokines (IL-1, TNF, and IL-6), IL-8,GROα, GROβ, GROγ or NAP-2 has the unique property of promotingneutrophil chemotaxis, enzyme release including but not limited toelastase release as well as superoxide production and activation. Theα-chemokines but particularly, GROα, GROβ, GROγ or NAP-2, workingthrough the IL-8 type I or II receptor can promote theneovascularization of tumors by promoting the directional growth ofendothelial cells. Therefore, the inhibition of IL-8 induced chemotaxisor activation would lead to a direct reduction in the neutrophilinfiltration.

Recent evidence also implicates the role of chemokines in the treatmentof HIV infections, Littleman et al., Nature 381, pp661 (1996) and Koupet al., Nature 381, pp 667 (1996).

The present invention also provides for a means of treating, in an acutesetting, as well as preventing, in those individuals deemed susceptibleto, CNS injuries by the chemokine receptor antagonist compounds ofFormula (I).

CNS injuries as defined herein include both open or penetrating headtrauma, such as by surgery, or a closed head trauma injury, such as byan injury to the head region. Also included within this definition isischemic stroke, particularly to the brain area.

Ischemic stroke may be defined as a focal neurologic disorder thatresults from insufficient blood supply to a particular brain area,usually as a consequence of an embolus, thrombi, or local atheromatousclosure of the blood vessel. The role of inflammatory cytokines in thisare has been emerging and the present invention provides a mean for thepotential treatment of these injuries. Relatively little treatment, foran acute injury such as these has been available.

TNF-α is a cytokine with proinflammatory actions, including endothelialleukocyte adhesion molecule expression. Leukocytes infiltrate intoischemic brain lesions and hence compounds which inhibit or decreaselevels of TNF would be useful for treatment of ischemic brain injury.See Liu et al., Stoke, Vol. 25., No. 7, pp 1481-88 (1994) whosedisclosure is incorporated herein by reference.

Models of closed head injuries and treatment with mixed 5-LO/CO agentsis discussed in Shohami et al., J. of Vaisc & Clinical Physiology andPharmacology, Vol. 3, No. 2, pp. 99-107 (1992) whose disclosure isincorporated herein by reference. Treatment which reduced edemaformation was found to improve functional outcome in those animalstreated.

The compounds of Formula (I) are administered in an amount sufficient toinhibit IL-8, binding to the IL-8 alpha or beta receptors, from bindingto these receptors, such as evidenced by a reduction in neutrophilchemotaxis and activation. The discovery that the compounds of Formula(I) are inhibitors of IL-8 binding is based upon the effects of thecompounds of Formulas (I) in the in vitro receptor binding assays whichare described herein. The compounds of Formula (1) have been shown, insome instances, to be dual inhibitors of both recombinant type I andtype II IL-8 receptors. Preferably the compounds are inhibitors of onlyone receptor, more preferably Type II.

As used herein, the term “IL-8 mediated disease or disease state” refersto any and all disease states in which IL-8, GROα, GROβ, GROγ, NAP-2 orENA-78 plays a role, either by production of IL-8, GROα, GROβ, GROγ,NAP-2 or ENA-78 themselves, or by IL-8, GROα, GROβ, GROγ, NAP-2 orENA-78 causing another monokine to be released, such as but not limitedto IL-1. IL-6 or TNF. A disease state in which, for instance, IL-1 is amajor component, and whose production or action, is exacerbated orsecreted in response to IL-8, would therefore be considered a diseasestated mediated by IL-8.

As used herein, the term “chemokine mediated disease or disease state”refers to any and all disease states in which a chemokine which binds toan IL-8 α or β receptor plays a role, such as but not limited IL-8,GROα, GROβ, GROγ, NAP-2 or ENA-78. This would include a disease state inwhich, IL-8 plays a role, either by production of IL-8 itself, or byIL-8 causing another monokine to be released, such as but not limited toIL-1, IL-6 or TNF. A disease state in which, for instance, IL-1is amajor component, and whose production or action, is exacerbated orsecreted in response to IL-8, would therefore be considered a diseasestated mediated by IL-8.

As used herein, the term “cytokine” refers to any secreted polypeptidethat affects the functions of cells and is a molecule which modulatesinteractions between cells in the immune, inflammatory or hematopoieticresponse. A cytokine includes, but is not limited to, monokines andlymphokines, regardless of which cells produce them. For instance, amonokine is generally referred to as being produced and secreted by amononuclear cell, such as a macrophage andlor monocyte. Many other cellshowever also produce monokines, such as natural killer cells,fibroblasts, basophils, neutrophils, endothelial cells, brainastrocytes, bone marrow stromal cells, epideral keratinocytes andB-lymphocytes. Lymphokines are generally referred to as being producedby lymphocyte cells. Examples of cytokines include, but are not limitedto, Interleukin-1 (IL-1), Interleukin-6 (IL-6), Interleukin-8 (IL-8),Tumor Necrosis Factor-alpha (TNF-α) and Tumor Necrosis Factor beta(TNF-β).

As used herein, the term “chemokine” refers to any secreted polypeptidethat affects the functions of cells and is a molecule which modulatesinteractions between cells in the immune, inflammatory or hematopoieticresponse, similar to the term “cytokine” above. A chemokine is primarilysecreted through cell transmembranes and causes chemotaxis andactivation of specific white blood cells and leukocytes, neutrophils,monocytes, macrophages, T-cells, B-cells, endothelial cells and smoothmuscle cells. Examples of chemokines include, but are not limited to,IL-8, GRO-α, GRO-β, GRO-γ, NAP-2, ENA-78, IP-10, MIP-1α, MIP-β, PF4, andMCP 1, 2, and 3.

In order to use a compound of Formula (I) or a pharmaceuticallyacceptable salt thereof in therapy, it will normally be formulated intoa pharmaceutical composition in accordance with standard pharmaceuticalpractice. This invention, therefore, also relates to a pharmaceuticalcomposition comprising an effective, non-toxic amount of a compound ofFormula (I) and a pharmaceutically acceptable carrier or diluent.

Compounds of Formula (I), pharmaceutically acceptable salts thereof andpharmaceutical compositions incorporating such may conveniently beadministered by any of the routes conventionally used for drugadministration, for instance, orally, topically, parenterally or byinhalation. The compounds of Formula (I) may be administered inconventional dosage forms prepared by combining a compound of Formula(I) with standard pharmaceutical carriers according to conventionalprocedures. The compounds of Formula (I) may also be administered inconventional dosages in combination with a known, second therapeuticallyactive compound. These procedures may involve mixing, granulating andcompressing or dissolving the ingredients as appropriate to the desiredpreparation. It will be appreciated that the form and character of thepharmaceutically acceptable character or diluent is dictated by theamount of active ingredient with which it is to be combined, the routeof administration and other well-known variables. The carrier(s) must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not deleterious to the recipient thereof.

The pharmaceutical carrier employed may be, for example, either a solidor liquid. Exemplary of solid carriers are lactose, terra alba, sucrose,talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acidand the like. Exemplary of liquid carriers are syrup, peanut oil, oliveoil, water and the like. Similarly, the carrier or diluent may includetime delay material well known to the arm, such as glycerylmono-stearate or glyceryl distearate alone or with a wax.

A wide variety of pharmaceutical forms can be employed. Thus, if a solidcarrier is used, the preparation can be tableted, placed in a hardgelatin capsule in powder or pellet form or in the form of a troche orlozenge. The amount of solid carrier will vary widely but preferablywill be from about 25 mg. to about 1 g. When a liquid carrier is used,the preparation will be in the form of a syrup, emulsion, soft gelatincapsule, sterile injectable liquid such as an ampule or nonaqueousliquid suspension.

Compounds of Formula (I) may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof Formula (I) externally to the epidermis or the buccal cavity and theinstillation of such a compound into the ear, eye and nose, such thatthe compound does not significantly enter the blood stream. In contrast,systemic administration refers to oral, intravenous, intraperitoneal andintramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as liniments, lotions, creams, ointmentsor pastes, and drops suitable for administration to the eye, ear ornose. The active ingredient may comprise, for topical administration,from 0.001% to 10% w/w, for instance from 1% to 2% by weight of theFormulation. It may however comprise as much as 10% w/w but preferablywill comprise less than 5% w/w, more preferably from 0. 1% to 1% w/w ofthe Formulation.

Lotions according to the present invention include those suitable forapplication to the skin or eye. An eye lotion may comprise a sterileaqueous solution optionally containing a bactericide and may be preparedby methods similar to those for the preparation of drops. Lotions orliniments for application to the skin may also include an agent tohasten drying and to cool the skin, such as an alcohol or acetone,and/or a moisturizer such as glycerol or an oil such as castor oil orarachis oil.

Creams, ointments or pastes according to the present invention aresemi-solid formulations of the active ingredient for externalapplication. They may be made by mixing the active ingredient infinely-divided or powdered form, alone or in solution or suspension inan aqueous or non-aqueous fluid, with the aid of suitable machinery,with a greasy or non-greasy base. The base may comprise hydrocarbonssuch as hard, soft or liquid paraffin, glycerol, beeswax, a metallicsoap; a mucilage; an oil of natural origin such as almond, corn,arachis, castor or olive oil; wool fat or its derivatives or a fattyacid such as steric or oleic acid together with an alcohol such aspropylene glycol or a macrogel. The formulation may incorporate anysuitable surface active agent such as an anionic, cationic or non-ionicsurfactant such as a sorbitan ester or a polyoxyethylene derivativethereof. Suspending agents such as natural gums, cellulose derivativesor inorganic materials such as silicaceous silicas, and otheringredients such as lanolin, may also be included.

Drops according to the present invention may comprise sterile aqueous oroily solutions or suspensions and may be prepared by dissolving theactive ingredient in a suitable aqueous solution of a bactericidaland/or fungicidal agent and/or any other suitable preservative, andpreferably including a surface active agent. The resulting solution maythen be clarified by filtration, transferred to a suitable containerwhich is then sealed and sterilized by autoclaving or maintaining at98-100 ° C. for half an hour. Alternatively, the solution may besterilized by filtration and transferred to the container by an aseptictechnique. Examples of bactericidal and fungicidal agents suitable forinclusion in the drops are phenylmercuric nitrate or acetate (0.002%),benzalkonium chloride (0.01%) and chlorhexidine acetate (0.01%).Suitable solvents for the preparation of an oily solution includeglycerol, diluted alcohol and propylene glycol.

Compounds of formula (I) may be administered parenterally, that is byintravenous, intramuscular, subcutaneous intranasal, intrarectal,intravaginal or intraperitoneal administration. The subcutaneous andintramuscular forms of parenteral administration are generallypreferred. Appropriate dosage forms for such administration may beprepared by conventional techniques. Compounds of Formula (I) may alsobe administered by inhalation, that is by intranasal and oral inhalationadministration. Appropriate dosage forms for such administration, suchas an aerosol formulation or a metered dose inhaler, may be prepared byconventional techniques.

For all methods of use disclosed herein for the compounds of Formula(I), the daily oral dosage regimen will preferably be from about 0.01 toabout 80 mg/kg of total body weight. The daily parenteral dosage regimenabout 0.001 to about 80 mg/kg of total body weight. The daily topicaldosage regimen will preferably be from 0.1 mg to 150 mg, administeredone to four, preferably two or three times daily. The daily inhalationdosage regimen will preferably be from about 0.01 mg/kg to about 1 mg/kgper day. It will also be recognized by one of skill in the art that theoptimal quantity and spacing of individual dosages of a compound ofFormula (I) or a pharmaceutically acceptable salt thereof will bedetermined by the nature and extent of the condition being treated, theform, route and site of administration, and the particular patient beingtreated, and that such optimums can be determined by conventionaltechniques. It will also be appreciated by one of skill in the art thatthe optimal course of treatment, i.e., the number of doses of a compoundof Formula (I) or a pharmaceutically acceptable salt thereof given perday for a defined number of days, can be ascertained by those skilled inthe art using conventional course of treatment determination tests.

The invention will now be described by reference to the followingbiological examples which are merely illustrative and are not to beconstrued as a limitation of the scope of the present invention.

BIOLOGICAL EXAMPLES

The IL-8, and Gro-α chemokine inhibitiory effects of compounds of thepresent invention were determined by the following in vitro assay:

Receptor Binding Assays

[¹²⁵I] IL-8 (human recombinant) was obtained from Amersham Corp.,Arlington Heights, Ill., with specific activity 2000 Ci/mmol. Gro-α wasobtained from NEN- New England Nuclear. All other chemicals were ofanalytical grade. High levels of recombinant human IL-8 type α and βreceptors were individually expressed in Chinese hamster ovary cells asdescribed previously (Holmes, et al., Science, 1991, 253, 1278). TheChinese hamster ovary membranes were homogenized according to apreviously described protocol (Haour, et aL, J Biol Chem., 249 pp2195-2205 (1974)). Except that the homogenization buffer was changed to10 mM Tris-HCL, 1 mM MgS04, 0.5 mM EDTA (ethylene-diaminetetra-aceticacid), 1 mMPMSF (α-toluenesulphonyl fluoride), 0.5 mg/L Leupeptin, pH7.5. Membrane protein concentration was determined using Pierce Co.micro-assay kit using bovine serum albumin as a standard. All assayswere performed in a 96-well micro plate format. Each reaction mixturecontained ¹²⁵I IL-8 (0.25 nM) or 125_(I) Gro-α and 0.5 μg/mL of IL-8Rαor 1.0 μg/mL of IL-8Rβ membranes in 20 MM Bis-Trispropane and 0.4 mMTris HCl buffers, pH 8.0, containing 1.2 mM MgSO_(4, 0.1) mM EDTA, 25 mMNaCl and 0.03% CHAPS. In addition, drug or compound of interest wasadded which had been pre-dissolved in DMSO so as to reach a finalconcentration of between 0.01 nM and 100 uM. The assay was initiated byaddition of ¹²⁵I-IL-8. After 1 hour at room temperature the plate washarvested using a Tomtec 96-well harvester onto a glass fiber filtermatblocked with 1% polyethylenimine/0.5% BSA and washed 3 times with 25 mMNaCl, 10 mM TrisHCl, 1 mM MgSO₄, 0.5 mM EDTA, 0.03% CHAPS, pH 7.4. Thefilter was then dried and counted on the Betaplate liquid scintillationcounter. The recombinant IL-8 Rα, or Type I, receptor is also referredto herein as the non-permissive receptor and the recombinant IL-8 Rβ, orType II, receptor is referred to as the permissive receptor.

All of the exemplified compounds of Formulas (I) noted herein in theSynthetic Chemistry Section, Example 1 to 15, demonstrated an IC₅₀ fromabout 45 to about <1 μg/mL in the permissive models for IL-8 receptorinhibition. Of those compounds tested, Examples 1 to 12 were also foundto be inhibitors of Gro-α binding at about the same level.

Chemotaxis Assay

The in vitro inhibitory properties of these compounds are determined inthe neutrophil chemotaxis assay as described in Current Protocols inImmunology, vol I, Suppl 1, Unit 6.12.3., whose disclosure isincorporated herein by reference in its entirety. Neutrophils whereisolated from human blood as described in Current Protocols inImmunology Vol I, Suppl 1 Unit 7.23.1, whose disclosure is incorporatedherein by reference in its entirety. The chemoattractants IL-8, GRO-α,GRO-β, GRO-γ and NAP-2 are placed in the bottom chamber of a 48multiwell chamber (Neuro Probe, Cabin John, Md.) at a concentrationbetween 0.1 and 100 nM. The two chambers are separated by a 5 umpolycarbonate filter. When compounds of this invention are tested, theyare mixed with the cells (0.001-1000 nM) just prior to the addition ofthe cells to the upper chamber. Incubation is allowed to proceed forbetween about 45 and 90 min at about 37° C. in a humidified incubatorwith 5% CO₂. At the end of the incubation period, the polycarbonatemembrane is removed and the top side washed, the membrane then stainedusing the Diff Quick staining protocol (Baxter Products, McGaw Park,Ill., USA). Cells which have chemotaxed to the chemokine are visuallycounted using a microscope. Generally, four fields are counted for eachsample, these numbers are averaged to give the average number of cellswhich had migrated. Each sample is tested in triplicate and eachcompound repeated at least four times. To certain cells (positivecontrol cells) no compound is added, these cells represent the maximumchemotactic response of the cells. In the case where a negative control(unstimulated) is desired, no chemokine is added to the bottom chamber.The difference between the positive control and the negative controlrepresents the chemotactic activity of the cells.

Elastase Release Assay

The compounds of this invention are tested for their ability to preventElastase release from human neutrophils. Neutrophils are isolated fromhuman blood as described in Current Protocols in Immunology Vol I, Suppl1 Unit 7.23.1. PMNs 0.88×10⁶ cells suspended in Ringer's Solution (NaCl118, KCl 4.56, NaHCO3 25, KH2PO4 1.03, Glucose 11.1, HEPES 5 mM, pH 7.4)are placed in each well of a 96 well plate in a volume of 50 ul. To thisplate is added the test compound (0.001-1000 nM) in a volume of 50 ul,Cytochalasin B in a volume of 50 ul (20 ug/ml) and Ringers buffer in avolume of 50 ul. These cells are allowed to warm (37° C., 5% CO2, 95%RH) for 5 min before IL-8, GROα, GROβ, GROγ or NAP-2 at a finalconcentration of 0.01-1000 nM was added. The reaction is allowed toproceed for 45 min before the 96 well plate is centrifuged (800×g 5 min)and 100 ul of the supernatant t re moved. This supernatant is added to asecond 96 well plate followed by an artificial elastase substrate(MeOSuc-Ala-Ala-Pro-Val-AMC, Nova Biochem, La Jolla, Calif.) to a finalconcentration of 6 ug/ml dissolved in phosphate buffered saline.Immediately, the plate is placed in a fluorescent 96 well plate reader(Cytofluor 2350, Millipore, Bedford, Mass.) and data collected at 3 minintervals according to the method of Nakajima et al J. Biol Chem 2544027 (1979). The amount of Elastase released from the PMNs is calculatedby measuring the rate of MeOSuc-Ala-Ala-Pro-Val-AMC degradation.

TNF-α in Traumatic Brain Injury Assay

The present assay provides for examination of the expression of tumornecrosis factor mRNA in specfic brain regions which followexperimentally induced lateral fluid-percussion traumatic brain injury(TBI) in rats. Adult Sprague-Dawley rats (n=42) were anesthetized withsodium pentobarbital (60 mg/kg, i.p.) and subjected to lateralfluid-percussion brain injury of moderate severity (2.4 atm.) centeredover the left temporaparietal cortex (n=18), or “sham” treatment(anesthesia and surgery without injury, n=18). Animals are sacrificed bydecapitation at 1,6 and 24 hr. post injury, brains removed, and tissuesamples of left (injured) parietal cortex (LC), corresponding area inthe contralateral right cortex (RC), cortex adjacent to injured parietalcortex (LA), corresponding adjacent area in the right cortex (RA), lefthippocampus (LH) and right hippocampus (RH) are prepared. Total RNA wasisolated and Northern blot hybridization is performed and quantitatedrelative to an TNF-α positive control RNA (macrophage=100%). A markedincrease of TNF-α mRNA expression is observed in LH (104±17% of positivecontrol, p<0.05 compared with sham), LC (105±21%, p<0.05) and LA (69±8%,p<0.01) in the traumatized hemisphere 1 hr. following injury. Anincreased TNF-α mRNA expression is also observed in LH (46±8%, p<0.05),LC (30±3%, p <0.01) and LA (32±3%, p<0.01) at 6 hr. which resolves by 24hr. following injury. In the contralateral hemisphere, expression ofTNF-α mRNA is increased in RH (46±2%, p<0.01), RC (4±3%) and RA (22±8%)at 1 hr. and in RH (28±11%), RC (7±5%) and RA (26±6%, p<f0.05) at 6 hr.but not at 24 hr. following injury. In sham (surgery without injury) ornaive animals, no consistent changes in expression of TNF-α mRNA areobserved in any of the 6 brain areas in either hemisphere at any times.These results indicate that following parasagittal fluid-percussionbrain injury, the temporal expression of TNF-α mRNA is altered inspecific brain regions, including those of the non-traumatizedhemisphere. Since TNF-α is able to induce nerve growth factor (NGF) andstimulate the release of other cytokines from activated astrocytes, thispost-traumatic alteration in gene expression of TNF-α plays an importantrole in both the acute and regenerative response to CNS trauma.

CNS Injury Model for IL-β mRNA

This assay characterizes the regional expression of interleukin- 1β(IL-1β) mRNA in specific brain regions following experimental lateralfluid-percussion traumatic brain injury (TBI) in rats. AdultSprague-Dawley rats (n=42) are anesthetized with sodium pentobarbital(60 mg/kg, i.p.) and subjected to lateral fluid-percussion brain injuryof moderate severity (2.4 atm.) centered over the left temporaparietalcortex (n=18), or “sham” treatment (anesthesia and surgery withoutinjury). Animals are sacrificed at 1, 6 and 24 hr. post injury, brainsremoved, and tissue samples of left (injured) parietal cortex (LC),corresponding area in the contralateral right cortex (RC), cortexadjacent to injured parietal cortex (LA), corresponding adjacent area inthe right cortex (RA), left hippocampus (LH) and right hippocampus (RH)are prepared. Total RNA is isolated and Northern blot hybridization wasperformed and the quantity of brain tissue 1β mRNA is presented aspercent relative radioactivity of IL-1β positive macrophage RNA whichwas loaded on same gel. At 1 hr. following brain injury, a marked andsignificant increase in expression of IL-1β mRNA is observed in LC(20.0±0.7% of positive control, n=6, p<0.05 compared with sham animal),LH (24.5±0.9%, p<0.05) and LA (21.5±3.1%, p<0.05) in the injuredhemisphere, which remained elevated up to 6 hr. post injury in the LC(4.0±%, n=6, p <0.05) and LH (5.0±1.3%, p<0.05). In sham or naiveanimals, no expression of IL-1β mRNA is observed in any of therespective brain areas. These results indicate that following TBI, thetemporal expression of IL-1β mRNA is regionally stimulated in specificbrain regions. These regional changes in cytokines, such as IL-1β play arole in the post-traumatic.

All publications, including but not limited to patents and patentapplications, cited in this specification are herein incorporated byreference as if each individual publication were specifically andindividually indicated to be incorporated by reference herein as thoughfully set forth.

The above description fully discloses the invention including preferredembodiments thereof. Modifications and improvements of the embodimentsspecifically disclosed herein are within the scope of the followingclaims. Without further elaboration, it is believed that one skilled inthe are can, using the preceding description, utilize the presentinvention to its fullest extent. Therefore the Examples herein are to beconstrued as merely illustrative and not a limitation of the scope ofthe present invention in any way. The embodiments of the invention inwhich an exclusive property or privilege is claimed are defined asfollows.

What is claimed is:
 1. A method of treating a chemokine mediated diseasestate, wherein the chemokine binds to an IL-8 α or β receptor in amammal, which comprises administering to said mammal an effective amountof a compound of the formula:

wherein X is oxygen or sulfur; R is hydroxy, carboxylic acid, thiol,SR₂, OR₂, —NH—C(O)R_(a), —C(O)NR₆R₇, —NHS(O)₂R_(b), —S(O)₂NHR_(c),NHC(X₂)NHR_(b), or tetrazolyl; wherein R₂ is a substituted aryl,heteroaryl, or heterocyclic ring which ring contains the functionalmoiety providing the ionizable hydrogen having a pKa of 10 or less;R_(a) is an alkyl, aryl, aryl C₁₋₄alkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, or a heterocyclic C₁₋₄alkyl moiety, all ofwhich may be optionally substituted; R_(b) is a NR₆R₇, alkyl, aryl,arylC₁₋₄alkyl, arylC₂₋₄alkenyl, heteroaryl, heteroarylC₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclic C₁₋₄alkyl,heterocyclic C₂₋₄alkenyl moiety, camphor, all of which may be optionallysubstituted one to three times independently by halogen; nitro;halosubstituted C₁₋₄ alkyl; C₁₋₄ alkyl; C₁₋₄ alkoxy; NR₉C(O)R_(a);C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; R₉ is hydrogen or a C₁₋₄ alkyl;R_(c) is alkyl, aryl, arylC₁₋₄alkyl, arylC₂₋₄alkenyl, heteroaryl,heteroarylC₁₋₄alkyl, heteroarylC₂₋₄alkenyl, heterocyclic, heterocyclicC₁₋₄alkyl, or a heterocyclic C₂₋₄alkenyl moiety, all of which may beoptionally substituted one to three times independently by halogen,nitro, halosubstituted C₁₋₄ alkyl, C₁₋₄ alkyl, C₁₋₄ alkoxy,NR₉C(O)R_(a), C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; and X₂ is oxygen orsulfur; R₁ is independently selected from hydrogen; halogen; nitro;cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)q S(O)_(t)R₄;hydroxy; hydroxy C₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; aryl C₁₋₄alkyloxy; heteroaryl; heteroarylalkyl; heterocyclic, heterocyclicC₁₋₄alkyl; heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀ alkenyl; heteroarylC₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenylC(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈;(CR₈R₈)q C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁(CR₈R₈)qC(O)OR₁₂; (CR₈R₈)q OC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)q NHS(O)₂R₁₇,(CR₈R₈)q S(O)₂NR₄R₅; or two R₁ moieties together may form O—(CH₂)_(s)O—or a 5 to 6 membered unsaturated ring; n is an integer having a value of1 to 3; m is an integer having a value of 1 to 3; q is 0, or an integerhaving a value of 1 to 10; t is 0, or an integer having a value of 1 or2; s is an integer having a value of 1 to 3; v is an integer having avalue of 1 to 4; R₄ and R₅ are independently hydrogen, optionallysubstituted C₁₋₄ alkyl, optionally substituted aryl, optionallysubstituted aryl C₁₋₄alkyl, optionally substituted heteroaryl,optionally substituted heteroaryl C₁₋₄alkyl, heterocyclic,heterocyclicC₁₋₄ alkyl, or R₄ and R₅ together with the nitrogen to whichthey are attached form a 5 to 7 member ring which may optionallycomprise an additional heteroatom selected from oxygen, nitrogen orsulfur; R₆ and R₇ are independently hydrogen or a C₁₋₄ alkyl group, orR₆ and R₇ together with the nitrogen to which they are attached form a 5to 7 member ring which ring may optionally contain an additionalheteroatom which heteroatom is selected from oxygen, nitrogen or sulfur;Y is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; S(O)_(t)R₄; hydroxy;hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; heterocyclic,heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; NR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅; C(O)NR₄R₅;C(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; C₁₋₁₀ alkyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁;C₂₋₁₀ alkenyl C(O)OR₁₁; C(O)R₁₁; C(O)OR₁₂; OC(O) R₁₁; NR₄C(O)R₁₁; or twoY moieties together may form O—(CH₂)_(s)O— or a 5 to 6 memberedunsaturated ring; R₈ is hydrogen or C₁₋₄ alkyl; R₁₀ is C₁₋₁₀ alkylC(O)₂R₈; R₁₁ is hydrogen, C₁₋₄ alkyl, optionally substituted aryl,optionally substituted aryl C₁₋₄alkyl, optionally substitutedheteroaryl, optionally substituted heteroarylC₁₋₄alkyl, optionallysubstituted heterocyclic, or optionally substitutedheterocyclicC₁₋₄alkyl; R₁₂ is hydrogen, C₁₋₁₀ alkyl, optionallysubstituted aryl or optionally substituted arylalkyl; R₁₃ and R₁₄ areindependently hydrogen, optionally substituted C₁₋₄ alkyl or one of R₁₃and R₁₄ may be optionally substituted aryl; R₁₇ is C₁₋₄alkyl, aryl,arylalkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, orheterocyclicC₁₋₄alkyl, wherein the aryl, heteroaryl and heterocyclicrings may all be optionally substituted; R_(d) is NR₆R₇, alkyl,arylC₁₋₄alklyl, arylC₂₋₄ alkenyl, heteroaryl, hetroaryl-C₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclicC₁₋₄ alkyl, whereinthe aryl, heteoaryl and heterocyclic rings may all be optionallysubstituted; provided that R₁ is not 3-phenyl, or 4-isopropyl, or5-phenyl or 5-ethylsulfonyl, when R is OH, v is 1, R₁₃ and R₁₄ arehydrogen, and Y is hydrogen; or a pharmaceutically acceptable saltthereof.
 2. The method according to claim 1 wherein the R₂ is optionallysubstituted one to three times by halogen, nitro, halosubstituted C₁₋₁₀alkyl, C₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, SH, —C(O)NR₆R₇,—NH—C(O)R_(a), —NHS(O)₂R_(b), S(O)NR₆R₇, C(O)OR₈, or a tetrazolyl ring.3. The method according to claim 1 wherein R is OH, —NHS(O)₂R_(b) orC(O)OH.
 4. The method according to claim 1 wherein R₁ is halogen, cyano,nitro, CF₃, C(O)NR₄R₅, alkenyl C(O)NR₄R₅, C(O) R₄R₁₀, alkenyl C(O)OR₁₂,heteroaryl, heteroarylalkyl, heteroaryl alkenyl, or S(O)NR₄R₅.
 5. Themethod according to claim 1 wherein Y is halogen, C₁₋₄ alkoxy,optionally substituted aryl, optionally substituted arylalkoxy,methylene dioxy, NR₄R₅, thioC₁₋₄alkyl, thioaryl, halosubstituted alkoxy,optionally substituted C₁₋₄alkyl, hydroxy alkyl.
 6. The method accordingto claim 1 wherein R is OH, SH, or NHS(O)₂R_(b) and R₁ is substituted inthe 3-position, the 4-position or di substituted in the 3,4-position byan electron withdrawing moiety.
 7. The method according to claim 1wherein the mammal is afflicted with a chemokine mediated diseaseselected from psoriasis, or atopic dermatitis, asthma, chronicobstructive pulmonary disease, adult respiratory distress syndrome,arthritis, inflammatory bowel disease, Crohn's disease, ulcerativecolitis, septic shock, endotoxic shock, gram negative sepsis, toxicshock syndrome, stroke, cardiac and renal reperfusion injury,glomerulo-nephritis, or thrombosis, alzheimers disease, graft vs. hostreaction, or allograft rejections.
 8. A compound of the formula:

wherein X is oxygen or sulfur; R is hydroxy, carboxylic acid, thiol,SR₂, OR₂, NH—C(O)R_(a), C(O)NR₆R₇, NHS(O)₂R_(b), S(O)₂NHR_(c),NHC(X₂)NHR_(b), or tetrazolyl; wherein R₂ is a substituted aryl,heteroaryl, or heterocyclic ring which ring contains the functionalmoiety providing the ionizable hydrogen having a pKa of 10 or less;R_(a) is an alkyl, aryl, aryl C₁₋₄alkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, or a heterocyclic C₁₋₄alkyl moiety, all ofwhich may be optionally substituted; R_(b) is a NR₆R₇, alkyl, aryl,arylC₁₋₄alkyl, arylC₂₋₄alkenyl, heteroaryl, heteroarylC₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclic C₁₋₄alkyl,heterocyclic C₂₋₄alkenyl moiety, camphor, all of which may be optionallysubstituted one to three times independently by halogen; nitro;halosubstituted C₁₋₄ alkyl; C₁₋₄ alkyl; C₁₋₄ alkoxy; NR₉C(O)R_(a);C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; R₉ is hydrogen or a C₁₋₄ alkyl;R_(c) is alkyl, aryl, arylC₁₋₄alkyl, arylC₂₋₄alkenyl, heteroaryl,heteroarylC₁₋₄alkyl, heteroarylC₂₋₄alkenyl, heterocyclic, heterocyclicC₁₋₄alkyl, or a heterocyclic C₂₋₄alkenyl moiety, all of which may beoptionally substituted one to three times independently by halogen,nitro, halosubstituted C₁₋₄ alkyl, C₁₋₄ alkyl, C₁₋₄ alkoxy,NR₉C(O)R_(a), C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; and X₂ is oxygen orsulfur; R₁ is independently selected from hydrogen; halogen; nitro;cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)q S(O)_(t)R₄;hydroxy; hydroxy C₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; aryl C₁₋₄alkyloxy; heteroaryl; heteroarylalkyl; heterocyclic, heterocyclicC₁₋₄alkyl; heteroaryl C₁₋₄ alkyloxy; aryl C₂₋₁₀ alkenyl; heteroarylC₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅; C₂₋₁₀ alkenylC(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈;(CR₈R₈)q C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁(CR₈R₈)qC(O)OR₁₂; (CR₈R₈)q OC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)q NHS(O)₂R₁₇,(CR₈R₈)q S(O)₂NR₄R₅; or two R₁ moieties together may form O—(CH₂)_(s)O—or a 5 to 6 membered unsaturated ring; n is an integer having a value of1 to 3; m is an integer having a value of 1 to 3; q is 0, or an integerhaving a value of 1 to 10; t is 0, or an integer having a value of 1 or2; s is an integer having a value of 1 to 3; v is an integer having avalue of 1 to 4; R₄ and R₅ are independently hydrogen, optionallysubstituted C₁₋₄ alkyl, optionally substituted aryl, optionallysubstituted aryl C₁₋₄alkyl, optionally substituted heteroaryl,optionally substituted heteroaryl C₁₋₄alkyl, heterocyclic,heterocyclicC₁₋₄ alkyl, or R₄ and R₅ together with the nitrogen to whichthey are attached form a 5 to 7 member ring which may optionallycomprise an additional heteroatom selected from oxygen, nitrogen orsulfur; R₆ and R₇ are independently hydrogen or a C₁₋₄ alkyl group, orR₆ and R₇ together with the nitrogen to which they are attached form a 5to 7 member ring which ring may optionally contain an additionalheteroatom which heteroatom is selected from oxygen, nitrogen or sulfur;Y is independently selected from hydrogen; halogen; nitro; cyano;halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀ alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy;halosubstituted C₁₋₁₀ alkoxy; azide; (CR₈R₈)q S(O)_(t)R₄; hydroxy;hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl; aryloxy; arylC₁₋₄ alkyloxy;heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄ alkyloxy; heterocyclic,heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl; heteroaryl C₂₋₁₀ alkenyl;heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)q NR₄R₅; C₂₋₁₀ alkenyl C(O)NR₄R₅;(CR₈R₈)q C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₁₀; S(O)₃H; S(O)₃R₈; (CR₈R₈)qC(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenyl C(O)OR₁₁; C(O)R₁₁;(CR₈R₈)q C(O)OR₁₂; (CR₈R₈)q OC(O) R₁₁; (CR₈R₈)q NR₄C(O)R₁₁, (CR₈R₈)qNHS(O)₂R_(d), (CR₈R₈)q S(O)₂NR₄R₅; or two Y moieties together may formO—(CH₂)_(s)O— or a 5 to 6 membered unsaturated ring; R₈ is hydrogen orC₁₋₄ alkyl; R₁₋₁₀ is C₁₋₁₀ alkyl C(O)₂R_(8;) R₁₁ is hydrogen, C₁₋₄alkyl, optionally substituted aryl, optionally substituted arylC₁₋₄alkyl, optionally substituted heteroaryl, optionally substitutedheteroarylC₁₋₄alkyl, optionally substituted heterocyclic, or optionallysubstituted heterocyclicC₁₋₄alkyl; R₁₂ is hydrogen, C₁₋₁₀ alkyl,optionally substituted aryl or optionally substituted arylalkyl; R₁₃ andR₁₄ are independently hydrogen, optionally substituted C₁₋₄ alkyl or oneof R₁₃ and R₁₄ may be optionally substituted aryl; R₁₇ is C₁₋₄alkyl,aryl, arylalkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, orheterocyclicC₁₋₄alkyl, wherein the aryl, heteroaryl and heterocyclicrings may all be optionally substituted; R_(d) is NR₆R₇, alkyl,arylC1-4alklyl, arylC₂₋₄ alkenyl, heteroaryl, hetroaryl-C₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclicC₁₋₄ alkyl, whereinthe aryl, heteoaryl and heterocyclic rings may all be optionallysubstituted; provided that R₁ is not 3-phenyl, or 4-isopropyl, or5-phenyl or 5-ethylsulfonyl, when R is OH, v is 1, R₁₃ and R₁₄ arehydrogen, and Y is hydrogen; or a pharmaceutically acceptable saltthereof.
 9. The compound according to claim 8 wherein the R₂ isoptionally substituted one to three times by halogen, nitro,halosubstituted C₁₋₁₀ alkyl, C₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, SH,—C(O)NR₆R₇, —NH—C(O)R_(a), —NHS(O)₂R_(b), S(O)NR₆R₇, C(O)OR₈, or atetrazolyl ring.
 10. The compound according to claim 8 wherein R is OH,—NHS(O)₂R_(b) or C(O)OH.
 11. The compound according to claim 8 whereinR₁ is halogen, cyano, nitro, CF₃, C(O)NR₄R₅, alkenyl C(O)NR₄R₅, C(O)R₄R₁₀, alkenyl C(O)OR₁₂, heteroaryl, heteroarylalkyl, heteroarylalkenyl, or S(O)NR₄R₅.
 12. The compound according to claim 8 wherein Yis halogen, C₁₋₄ alkoxy, optionally substituted aryl, optionallysubstituted arylalkoxy, methylene dioxy, NR₄R₅, thioC₁₋₄alkyl, thioaryl,halosubstituted alkoxy, optionally substituted C₁₋₄alkyl, hydroxy alkyl.13. The compound according to claim 8 wherein R is OH, SH, orNHS(O)₂R_(b) and R₁ is substituted in the 3-position, the 4-position ordi substituted in the 3,4-position by an electron withdrawing moiety.14. A pharmaceutical composition comprising an effective amount of acompound according to claim 8, and a phamaceutically acceptable carrieror diluent.
 15. A compound of the formula:

wherein X is oxygen or sulfur; R is any functional moiety having anionizable hydrogen and a pKa of 10 or less; R₁ is independently selectedfrom hydrogen; halogen; nitro; cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide;(CR₈R₈)q S(O)_(t)R₄; hydroxy; hydroxy C₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl;aryloxy; aryl C₁₋₄ alkyloxy; heteroaryl; heteroarylalkyl; heterocyclic,heterocyclic C₁₋₄alkyl; heteroaryl C₁₋₄ alkloxy; aryl C₂₋₁₀ alkenyl;heteroaryl C₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)qNR₄R₅;C₂₋₁₀ alkenyl C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₁₀;S(O)₃H; S(O)₃R₈; (CR₈R₈)q C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenylC(O)OR₁₁ (CR₈R₈)q C(O)OR₁₂; (CR₈R₈)q OC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁,(CR₈R₈)q NHS(O)₂R₁₇, (CR₈R₈)q S(O)₂NR₄R₅; or two R₁ moieties togethermay form O—(CH₂)_(s)O— or a 5 to 6 membered unsaturated ring; n is aninteger having a value of 1 to 3; m is an integer having a value of 1 to3; q is 0, or an integer having a value of 1 to 10; t is 0, or aninteger having a value of 1 or 2; s is an integer having a value of 1 to3; v is an integer having a value of 1 to 4; R₄ and R₅ are independentlyhydrogen, optionally substituted C₁₋₄ alkyl, optionally substitutedaryl, optionally substituted aryl C₁₋₄alkyl, optionally substitutedheteroaryl, optionally substituted heteroaryl C₁₋₄alkyl, heterocyclic,heterocyclic C₁₋₄ alkyl, or R₄ and R₅ together with the nitrogen towhich they are attached form a 5 to 7 member ring which may optionallycomprise an additional heteroatom selected from O/N/S; R₆ and R₇ areindependently hydrogen or a C₁₋₄ alkyl group, or R₆ and R₇ together withthe nitrogen to which they are attached form a 5 to 7 member ring whichring may optionally contain an additional heteroatom which heteroatom isselected from oxygen, nitrogen or sulfur; Y is independently selectedfrom hydrogen; halogen; nitro; cyano; halosubstituted C₁₋₁₀ alkyl; C₁₋₁₀alkyl; C₂₋₁₀ alkenyl; C₁₋₁₀ alkoxy; halosubstituted C₁₋₁₀ alkoxy; azide;(CR₈R₈)q S(O)_(t)R₄; hydroxy; hydroxyC₁₋₄alkyl; aryl; aryl C₁₋₄ alkyl;aryloxy; arylC₁₋₄ alkyloxy; heteroaryl; heteroarylalkyl; heteroaryl C₁₋₄alkyloxy; heterocyclic, heterocyclic C₁₋₄alkyl; aryl C₂₋₁₀ alkenyl;heteroaryl C₂₋₁₀ alkenyl; heterocyclic C₂₋₁₀ alkenyl; (CR₈R₈)q NR₄R₅;C₂₋₁₀ alkenyl C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₅; (CR₈R₈)q C(O)NR₄R₁₀;S(O)₃H; S(O)₃R₈; (CR₈R₈)q C(O)R₁₁; C₂₋₁₀ alkenyl C(O)R₁₁; C₂₋₁₀ alkenylC(O)OR₁₁; C(O)R₁₁; (CR₈R₈)q C(O)OR₁₂; (CR₈R₈)q OC(O) R₁₁; (CR₈R₈)qNR₄C(O)R₁₁, (CR₈R₈)q NHS(O)₂R_(d), (CR₈R₈)q S(O)₂NR₄R₅; or two Ymoieties together may form O—(CH₂)_(s)O— or a 5 to 6 memberedunsaturated ring; R₈ is hydrogen or C₁₋₄ alkyl; R₁₀ is C₁₋₁₀ alkylC(O)₂R₈; R₁₁ is hydrogen, C₁₋₄ alkyl, optionally substituted aryl,optionally substituted aryl C₁₋₄alkyl, optionally substitutedheteroaryl, optionally substituted heteroarylC₁₋₄alkyl, optionallysubstituted heterocyclic, or optionally substitutedheterocyclicC₁₋₄alkyl; R₁₂ is hydrogen, C₁₋₁₀ alkyl, optionallysubstituted aryl or optionally substituted arylalkyl; R₁₃ and R₁₄ areindependently hydrogen, optionally substituted C₁₋₄ alkyl, or one of R₁₃and R₁₄ may be optionally substituted aryl; R₁₇ is C₁₋₄alkyl, aryl,arylalkyl, heteroaryl, heteroarylC₁₋₄alkyl, heterocyclic, orheterocyclicC₁₋₄alkyl, wherein the aryl, heteroaryl and heterocyclicrings may all be optionally substituted; R_(d) is NR₆R₇, alkyl,arylC1-4alklyl, arylC₂₋₄ alkenyl, heteroaryl, hetroaryl-C₁₋₄alkyl,heteroarylC₂₋₄ alkenyl, heterocyclic, heterocyclicC₁₋₄ alkyl, whereinthe aryl, heteoaryl and heterocyclic rings may all be optionallysubstituted; E is optionally selected from

 the asterix * denoting point of attachment of the ring; and Z isoptionally selected from

 the asterix * denoting point of attachment of the ring, with at leastone E ring or Z ring being present; or a pharmaceutically acceptablysalt thereof.
 16. The compound according to claim 15 wherein theionizable hydrogen has a pKa of 3 to
 10. 17. The method according toclaim 15 wherein R is hydroxy, carboxylic acid, thiol, SR₂, OR₂,NH—C(O)R_(a), C(O)NR₆R₇, NHS(O)₂R_(b), S(O)₂NHR_(c), NHC(X₂)NHR_(b), ortetrazolyl; wherein R₂ is a substituted aryl, heteroaryl, orheterocyclic ring which ring contains the functional moiety providingthe ionizable hydrogen having a pKa of 10 or less; R_(a) is an alkyl,aryl, aryl C₁₋₄alkyl, heteroaryl, heteroaryl C₁₋₄alkyl, heterocyclic, ora heterocyclic C₁₋₄alkyl moiety, all of which may be optionallysubstituted; R_(b) is a NR₆R₇, alkyl, aryl, arylC₁₋₄alkyl,arylC₂₋₄akenyl, heteroaryl, heteroarylC₁₋₄alkyl, heteroarylC₂₋₄ alkenyl,heterocyclic, heterocyclic C₁₋₄alkyl, heterocyclic C₂₋₄alkenyl moiety,camphor, all of which may be optionally substituted one to three timesindependently by halogen; nitro; halosubstituted C₁₋₄ alkyl; C₁₋₄ alkyl;C₁₋₄ alkoxy; NR₉C(O)R_(a); C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; R₉ ishydrogen or a C₁₋₄ alkyl; R_(c) is alkyl, aryl, arylC₁₋₄alkyl,arylC₂₋₄alkenyl, heteroaryl, heteroarylC₁₋₄alkyl, heteroarylC₂₋₄alkenyl,heterocyclic, heterocyclic C₁₋₄alkyl, or a heterocyclic C₂₋₄alkenylmoiety, all of which may be optionally substituted one to three timesindependently by halogen, nitro, halosubstituted C₁₋₄ alkyl, C₁₋₄ alkyl,C₁₋₄ alkoxy, NR₉C(O)R_(a), C(O)NR₆R₇, S(O)₃H, or C(O)OC₁₋₄ alkyl; and X₂is oxygen or sulfur.
 18. The compound according to claim 17 wherein theR₂ is optionally substituted one to three times by halogen, nitro,halosubstituted C₁₋₁₀ alkyl, C₁₋₁₀ alkyl, C₁₋₁₀ alkoxy, hydroxy, SH,C(O)NR₆R₇, NH—C(O)R_(a), NHS(O)₂R_(b), S(O)NR₆R₇, C(O)OR₈, or atetrazolyl ring.
 19. The compound according to claim 17 wherein R is OH,NHS(O)₂R_(b) or C(O)OH.
 20. The compound according to claim 15 whereinR₁ is halogen, cyano, nitro, CF₃, C(O)NR₄R₅, alkenyl C(O)NR₄R₅, C(O)R₄R₁₀, alkenyl C(O)OR₁₂, heteroaryl, heteroarylalyl , heteroarylalkenyl, or S(O)NR₄R₅.
 21. The compound according to claim 15 wherein Yis halogen, C₁₋₄ alkoxy, optionally substituted aryl, optionallysubstituted arylalkoxy, methylene dioxy, NR₄R₅, thioC₁₋₄alkyl, thioaryl,halosubstituted alkoxy, optionally substituted C₁₋₄alkyl, or hydroxyalkyl.
 22. The compound according to claim 15 wherein R is OH, SH, orNHS(O)₂R_(b) and R₁ is substituted in the 3-position, the 4-position ordi substituted in the 3,4-position by an electron withdrawing moiety.23. A pharmaceutical composition comprising an effective amount of acompound according to claim 15, and a pharmaceutically acceptablecarrier or diluent.